Discrete acyltransferases associated with type I polyketide synthases and methods of use

ABSTRACT

Genetic and biochemical characterization of the leinamycin biosynthesis gene cluster from  Streptomyces atroolivaceus  S-140 revealed two PKS genes, lnmI and lnmJ, that encode six PKS modules, none of which contains a cognate AT domain. The AT activity is provided in trans by a discrete protein, LnmG, which loads the malonyl coenzyme A extender unit onto the ACP domains of all six PKS modules. This finding provides a basis for methods of engineering modular polyketide synthases and polyketide synthase/nonribosomal peptide synthetases.

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This is a continuation-in-part application of InternationalApplication PCT/US02/08937, filed on Mar. 22, 2002 which claimed thebenefit of and priority to U.S. Provisional Application No. 60/278,935,filed on Mar. 26, 2001, both of which are incorporated herein byreference in their entirety for all purposes.

STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSOREDRESEARCH AND DEVELOPMENT

[0002] This work was supported in part by grants from the NationalInstitutes of Health (NIH)-CA 78747 and the National Science Foundation(NSF)-MCB 0196528. The Government of the United States of America mayhave certain rights in this invention.

FIELD OF INVENTION

[0003] This invention relates to the field of polyketide synthesis. Inparticular, this invention pertains to type I polyketide synthasesrequiring discrete acyltransferases and methods of using the discreteacyltransferases for the production of novel products.

BACKGROUND OF THE INVENTION

[0004] Polyketides and nonribosomal peptides are two large families ofnatural products that include many clinically valuable drugs, such aserthromycin and vancomycin (antibacterial), FK506 and cyclosporin(immunosuppressant), and epothilone, and bleomycin, or leinamycin(antitumor). The biosyntheses of polyketides and nonribosomal peptidesare catalyzed by polyketide synthases (PKSs) (Hopwood (1997) Chem. Rev.,97: 2465; Katz (1997) Chem. Rev., 97: 22557; C Khosla, (1997) Chem.Rev., 97: 22577; Ikeda and Omura, (1997) Chem. Rev., 97: 2591; Stauntonand Wilkinson (1997) Chem. Rev., 97: 2611; Cane et al. (1998) Science282: 63) and nonribosomal peptide synthetases (NRPSs) Cane et al. (1998)Science 282: 63). Marahiel et al. (1997) Chem. Rev., 97: 2651; vonDöhren et al. (1997) Chem. Rev., 97: 2675), respectively. Remarkably,PKSs and NRPSs use a very similar strategy for the assembly of these twodistinct classes of natural products by sequential condensation of shortcarboxylic amino acids, respectively, and utilize the same4′-phosphopantetheine prosthetic group, via a thioster linkage, tochannel the growing polyketide or peptide intermediate during theelongation processes.

[0005] Both type I PKSs and NRPSs are multifunctional proteins that areorganized into modules. A module is defined as a set of distinctivedomains that encode all the enzyme activities necessary for one cycle ofpolyketide or peptide chain elongation and associated modifications. Thenumber and order of modules and the type of domains within a module oneach PKS or NRPS protein determine the structural variations of theresulting polyketide and peptide products by dictating the number,order, choice of the carboxylic acid or amino acid to be incorporated,and the modifications associated with a particular cycle of elongation.Since the modular architecture of both PKS (Cane et al. (1998) Science,282: 63; Katz and Danadio (1993) Ann. Rev. Microbiol. 47:875 (1993);Hutchinson and Fuji (1995) Ann. Rev. Microbiol. 49: 201) and NRPS (Caneet al. (1998) Science 282: 63, Stachelhaus et al. (1995) Science 269:69, Stachelhaus et al. (1998) Mol. Gen. Genet. 257: 308; Belshaw et al.(1999) Science 284: 486) has been exploited successfully incombinatorial biosynthesis of diverse “unnatural” natural products, ahybrid PKS and NRPS system, capable of incorporating both caroboxylicacids and amino acids into the final products, can lead to even greaterchemical structural diversity.

[0006] Leinamycin (Lnm) is a novel antitumor antibiotic produced byseveral Streptomyces species (Hara et al. (1989) J. Antiobiot. 42:333-335; Hara et al. (1989) J. Antiobiot. 42: 1768-1774; Nakano et al.(1992) Pages 72-75 In Harnessing Biotechnol. 21^(st) Center, Proc. Int.Biotechnol. Symp. Expo. 9^(th), Ladisch, M. R. and Bose, A., eds., ACS:Washington, D.C.). Its structure was revealed by X-raycrystallographical (Hirayama and Matsuzawa (1993) Chem. Lett.(1957-1958) and spectroscopic analyses (Hara et al. (1989) J. Antiobiot.42: 333-335; Hara et al. (1989) J. Antiobiot. 42: 1768-1774; andconfirmed by total synthesis (Kandra and Fukuyama (1993) J. Am. Chem.Soc. 115:8451-8452; Fukuyama and Kanda (1994) J. Synth. Org. Chem.Japan, 52: 888-899). It contains an unusual 1,3-dioxo-1,2-dithiolanemoiety that is spiro-fused to thiazole-containing 18-membered lactamring, a molecular architecture that has not been found to date in anyother natural product (FIG. 1).

[0007] Lnm exhibits a broad spectrum of antimicrobial activity againstGram-positive and Gram-negative bacteria, but not against fungi. Lnmshows potent antitumor activity in murine tumor models in vivo,including HELA S3, sarcoma 180, B-16, Colon 26, and leukemia P388. It isalso active against murine models inoculated with tumors that areresistant to clinically important antitumor drugs, such as cisplatin,doxorubicin, mitomycin, or cyclophosphamide (Hara et al. (1989) J.Antibiot. 42: 333-335; Hara et al. (1989) J. Antiobiot. 42: 1768-1774;Nakano et al. In Harnessing Biotechnol. 21^(st) Century, Proc. Int.Biotechnol. Symp. Expo. 9^(th), Ladish, M. R. and Bose, A., eds., ACS:Washington, D.C.). Lnm preferentially inhibits DNA synthesis andinteracts directly with DNA to cause single-strand scission of DNA inthe presence of thiol agents as cofactors. The presence of the sulfoxidegroup in the dithiolane moeity is essential for the DNA-cleavingactivity Hara et al. (1990) Biochemistry 29: 5676-5681). Interestingly,simple 1,3-dioxo-1,2-dithioilanes are also thiol-dependent DNA cleavingagents in vitro (Behroozi et al. (1995) J. Org. Chem. 60: 3964-3966;Behroozi et al. (1996) Biochemistry 35: 1768-1774; Mitra et al. (1997)J. Am. Chem. Soc. 119: 11691-11692). However, the mechanisms for DNAcleavage by simple 1,3-dioxo-1,2-dithiolanes and Lnm are distinctoxidative cleavage by 1,3-dioxo-1,2-dithiolanes that convert molecularoxygen to DNA-cleaving oxygen radicals mediated by polysulfides(Behroozi et al. (1995) J. Org. Chem. 60: 3964-3966; Behroozi et al.(1996) Biochemistry 35: 1768-1774; Mitra et al. (1997) J. Am. Chem. Soc.119: 11691-11692) and alkylative cleavage by Lnm mediated by anepisulfonium ion intermediate (Mitra et al. (1997) J. Chem. Soc. 119:11691-11692; Asai et al. (1996) J. Am. Chem. Soc. 118:6802-6803; Asai etal. (1997) Bioorg. Med. Chem. 5: 723-729) (FIG. 1). The latter mechanismrepresents an unprecedented mode of action for the thiol-dependent DNAcleavage by Lnm.

[0008] Aimed at discovering clinically useful Lnm analogs, both totalsynthesis (Kandra and Fukuyama (1993) J. Chem. Soc. 115: 8451-8452;Fukuyama and Kandra (1994) J. Synth. Org. Chem. Japan, 52: 888-899;Pattenden and Shuker (1991) Tetrahedron Lett. 32:6625-6628; Pattendenand Shuker (1992) J. Chem. Soc. Perkin Trans I, 1215-1221; Kandra et al.(1992) Tetrahedron Lett. 33: 5701-5704; Pattenden and Thom (1993)Synlett 215-216) and chemical modification of the natural Lnm have beeninvestigated. Modifications at both C-8 hydroxy and C-9 keto groups aswell as the 1,3-dioxo-1,2-dithiolane moiety have generated a number ofLnm analogs with improved antitumor activity and in vivo stability(Kandra et al. (1998) Bioorg. Med. Chem. Lett. 8: 909-912; Kandra et al.(1999) J. Med. Chem. 42: 1330-1332), supporting the wisdom of makingnovel anticancer drugs based on the Lnm scaffold. However, for a complexmolecule like Lnm, chemical total synthesis has very limited practicalvalue, and chemical modification only can access to limited functionalgroups, often requiring multiple extra protection/deprotection steps.

SUMMARY OF THE INVENTION

[0009] This invention pertains to the isolation and elucidation of theleinamycin gene cluster (see SEQ ID NO:1). This gene cluster (nucleicacid sequence) encodes all of the open reading frames (ORFs) that encodepolypeptides sufficient to direct the biosynthesis of leinamycin. Thenucleic acids can be used in their “native” format or recombined in awide variety of manners to create novel synthetic pathways.

[0010] Thus, in one embodiment, this invention provides an isolatednucleic acid comprising a nucleic acid selected from the groupconsisting of: 1) A nucleic acid encoding one or more leinamycin (lnm)open reading frames (ORFs) identified in Tables 1 and 2(ORFs—35 through−1, lnmA through lnmZ, and +1 through +9)). A nucleic acid encoding apolypeptide encoded by any one or more of leinamycin (lnm) open readingframes (ORFs) identified in Tables 1 and 2 (ORFs−35 through−1, lnmAthrough lnmZ, and +1 through +9); 3) A nucleic acid comprising thenucleotide sequence of a nucleic acid amplified by polymerase chainreaction (PCR) using any one of the primer pairs identified in Table 2and the nucleic acid of a leinamycin-producing organism as a template;and 4) A nucleic acid that encodes a protein comprising at least onecatalytic domain selected from the group consisting of a condensation(c) domain, an adenylation (A) domain, a peptidyl carrier protein (PCP)domain, a condensation/cyclization (cy), an acyl-carrier protein(ACP)-like domain, an oxidization domain (Ox), and enoyl reductasedomain (ER), a methyltransferase domain, a phosphotransferase domain, apeptide synthase domain, and an aminotransferase domain, and thatspecifically hybridizes to on or more of lnm ORFs −35 through −1, lnmAthrough lnmZ and/or +1 through +9 under stringent conditions.

[0011] In certain embodiments, the isolated nucleic acid comprises anucleic acid encoding at least two, preferably at least three, morepreferably at least four, and most preferably at least five, openreading frames independently selected from the group consisting ofleinamycin (lnm) open reading frames −35 through −1, lnmA through lnmZand +1 through +9. In particularly preferred embodiments, the isolatednucleic acid encodes a module. In another embodiment, the nucleic acidcomprises a nucleic acid encoding a module comprising two or morecatalytic domains of a protein encoded by a nucleic acid of a leinamycin(lnm) gene cluster where the catalytic domains are selected from groupconsisting of a condensation (C) domain, an adenylation (A) domain, apeptidyl-carrier protein (PCP) domain, a condensation/cyclization domain(Cy), an acyl-carrier protein (ACP)-like domain, an oxidation domain(Ox), an enoyl reductase domain, a methyltransferase domain, aphosophotransferase domain, a peptide synthetase domain, and anaminotransferase domain. In particularly preferred embodiments, thenucleic acid comprises an open reading frame from SEQ ID NO: 1 or thecomplement thereof (e.g. as described in Table 2). In other preferredembodiments, the nucleic acid has the nucleotide sequence of a nucleicacid amplified by polymerase chain reaction (PCR) using any one of theprimer pairs identified in Table 2 and the nucleic acid of aleinamyicin-producing organism (e.g. S. atroolivaceus) as a template.

[0012] In still yet another embodiment, this intervention provides anisolated nucleic acid comprising a leinamycin (lnm) open reading frame(ORF) or an allelic variant thereof (e.g. a single nucleotidepolymorphism (SNP) of a leinamycin (lnm) open reading frame (ORF)).

[0013] In another embodiment, this invention provides an isolated genecluster comprising open reading frames encoding polypeptides sufficientto direct the assembly of a leinamycin.

[0014] In one embodiment, this invention provides an isolated nucleicacid encoding a multi-functional protein complex comprising both apolyketide synthase (PKS) and a peptide synthetase (NRPS), where thepolyketide synthase or the peptide synthetase, has the amino acidsequence of a PKS or an NRPS encoded by the leinamycin (lnm) genecluster.

[0015] This invention also provides for various proteins. Thus, in oneembodiment, this invention provides an isolated multi-functional proteincomplex comprising both a polyketide synthase (PKS) and a peptidesynthetase (NRPS), where the polyketide synthase (PKS) and/or thepeptide synthetase (NRPS) has the amino acid sequence of a PKS or anNRPS found encoded by a nucleic acid from the leinamycin gene cluster.

[0016] In another embodiment, this invention provides a polypeptideselected from the group consisting of: 1) A catalytic domain encoded byone or more leinamycin (lnm) open reading frames (ORFs) e.g. ORFs −35through −1, lnmA through lnmZ, and +1 through +9, (e.g. as identified inTable 2); 2) A catalytic domain encoded by a nucleic acid having thesequence of a nucleic acid amplified by polymerase chain reaction (PCR)using any one of the primer pairs identified Table 2; and 3) A modulecomprising two or more catalytic domains of a protein encoded by anucleic acid of a leinamycin gene cluster. In preferred embodiments, thepolypeptide comprises an enzymatic domain selected from the groupconsisting of a condensation (C) domain, an adenylation (A) domain, apeptidyl carrier protein (PCP) domain, a condensation/cyclization domain(Cy), an acyl-carrier protein (ACP)-like domain, an oxidization domain(Ox), an NADH dehydrogenase domain, a methyltransferase domain, aphosphotransferase domain, a peptide synthetase domain, and anaminotransferase domain. In certain embodiments, the polypeptidecomprises domains encoded by at least two, preferably at least three,more preferably at least four, and most preferably at least five, openreading frames independently selected from the group consisting ofleinamycin (lnm) open reading frames 1 through 72. In certainembodiments, the polypeptide can comprise a module comprising two ormore catalytic domains of a protein encoded by a nucleic acid of aleinamycin gene cluster where the catalytic domains are selected fromthe group consisting of a condensation (C) domain, an adenylation(A)domain, a peptidyl carrier protein (PCP) domain, acondensation/cyclization domain (Cy), an acyl-carrier protein (ACP)-likedomain, an oxidization domain (Ox), an enoyl reductase domain (ER), amethyltransferase domain, a phosphotransferase domain, a peptidesynthetase domain, and an aminotransferase domain.

[0017] In another embodiment this invention provides an isolatedpolypeptide comprising a module where the module is specifically boundby an antibody that specifically binds to a leinamycin (lnm) module(e.g. is cross-reactive with an lnm polypeptide). In preferredembodiments, the polypeptide is specifically bound by an antibody thatspecifically binds to a polypeptide encoded by a leinamycin openingreading frame.

[0018] In certain embodiments, this invention provides an expressionvector comprising any one or more of the nucleic acids described herein.The nucleic acids are preferably operably linked to a promoter (e.g.constitutive promoter, inducible promoter, tissue specific promoter,etc.). Also provided are host cells transfected and/or transformed withsuch a vector. Thus, in one embodiment this invention provides a hostcell (e.g. bacterial cell) transfected and/or transformed with anexogenous nucleic acid comprising a gene cluster encoding polypeptidessufficient to direct the assembly of a leinamycin or leinamycin analogand/or with a nucleic acid sufficient to introduce a modification into alnm gene cluster (e.g. via homologous recombination). Particularlypreferred cells include, but are not limited to, eukaryotic cells,insect cells, and bacterial cells (e.g. Streptomyces cells).

[0019] This invention also provides a method of chemically modifying amolecule. The method involves contacting a molecule that is a substratefor a polypeptide encoded by one or more leinamycin biosynthesis genecluster open reading frames (e.g. a leinamycin intermediate metabolite)with a polypeptide encoded by one or more leinamycin biosynthesis genecluster open reading frames, whereby the polypeptide chemically modifiesthe molecule. In preferred embodiments, the method comprises contactingthe molecule with at least two, preferably at least 3, more preferablyat least 4 and most preferably at least 5 different polypeptides encodedby leinamycin (lnm) gene cluster open reading frames. The contacting canbe ex vivo or in a host cell (e.g. a bacterium such as Streptomyces).The molecule can include, but is not limited to, an endogenousmetabolite produced by the host cell or an exogenous suppliedmetabolite. In certain embodiments, the cell is a eukaryotic cell (e.g.a mammalian cell, a yeast cell, a plant cell, a fungal cell, and aninsect cell). In certain embodiments, the (substrate) molecule is anamino acid and the polypeptide is a peptide synthetase. In certainembodiments, the polypeptide is an amino transferase.

[0020] In still another embodiment, this invention provides a cell thatoverexpresses leinamycin. A particularly preferred cell overexpresses apolypeptide encoded by leinamycin open reading frame lnmG and/or lnmL.

[0021] This invention also provides a method of coupling a first aminoacid to a second amino acid. The method involves comprising contactingthe first and second amino acid with a recombinantly expressedleinamycin nonribosomal peptide synthetase (NRPS) (e.g. NRPS-1, NRPS-2,etc.). The contacting can be ex vivo or in a host cell (e.g. abacterium).

[0022] This invention also provides a method f coupling a first fattyacid to a second fatty acid. This method involves contacting the firstand second fatty acids with a recombinantly expressed leinamycinpolyketide synthase (PKS) (e.g. PKS-1, PKS-2, PKS-3, PKS-4, PKS-5,PKS-6). The contacting can be ex vivo or in a host cell (e.g. abacterium).

[0023] In one embodiment, this invention provides a method of producinga leinamycin or leinamycin analog. The method involves providing a celltransformed with an exogenous nucleic acid comprising a leinamycin genecluster encoding polypeptides sufficient to direct the assembly of theleinamycin or leinamycin analog; culturing the cell under conditionspermitting the biosynthesis of leinamycin or leinamycin analog; andisolating the leinamycin or leinamycin analog from the cell.

[0024] In another embodiment, this invention provides a method ofproducing a leinamycin analog. The method involves providing a cellcomprising a leinamycin gene cluster, transfecting the cell with anucleic acid that alters the leinamycin gene cluster through homologousrecombination, culturing the cell under conditions permitting thebiosynthesis of the leinamycin analog; and isolating the leinamycinanalog from the cell.

[0025] In still another embodiment, this invention provides an isolatednucleic acid comprising a nucleic acid encoding a phosphopantetheinyltransferase (PPTase) the nucleic acid encoding a phosphopantetheinyltransferase being selected from the group consisting of: 1) a nucleicacid encoding the protein comprising the amino acid sequence encoded bysap (Streptomyces altroolivaceus phosphopantetheinyl transferase) ofFIG. 5; 2) a nucleic acid encoding a polypeptide havingphosphopantetheinyl transferase activity where said nucleic acidspecifically hybridizes to the nucleic acid having the sequence encodedby sap (Streptomyces altroolivaceus phosphopantetheinyl transferase) ofFIG. 5 under stringent conditions. In a particularly preferredembodiment, the nucleic acid comprises the sequence of lmp in FIG. 5. Asdescribed above, the nucleic acid comprises a vector and cellscomprising such a vector are provided herein. Also provided is apolypeptide encoded by a phosphopantetheinyl transferase nucleic aciddescribed herein.

[0026] This invention also provides a method of converting anapo-carrier protein to a holo-carrier protein comprising reacting theapo-carrier protein with a recombinant phosphopantetheinyl transferaseencoded by the lnm PPTase nucleic acid described herein and coenzyme Athereby producing a holo-carrier protein.

[0027] In still yet another embodiment, this invention provides a cellcomprising a modified leinamycin gene cluster nucleic acid, where thecell produces elevated amounts of leinamycin as compared to the wildtype cell. Particularly preferred cells overexpress a resistance genefrom the leinamycin gene cluster (e.g. a resistance gene listed in Table2).

[0028] This invention also provides antibodies that specifically bind toa polypeptide encoded by an lnm open reading frame identified in Table2. The antibodies include, but are not limited to intact antibodies,antibody fragments, and single chain antibodies.

[0029] Based upon the present inventors' discovery that the leinamycinbiosynthesis gene cluster utilizes a discrete AT, LnmG, to provideacyltransferase activity in trans to a type I polyketide synthaselacking modular AT domains, the invention is further directed to methodsexploiting this unique and useful observation. For example, theinvention is further directed to methods of loading an extender moleculeon an acyl carrier protein (ACP) domain including the step of contactingthe ACP domain with the extender molecule and a recombinantly expressed,discrete acyl transferase (AT). The extender molecule is loaded onto theACP domain by the catalytic activity of the discrete AT, provided intrans. The method may be carried out in vitro or in a host cell. If in ahost cell, the host cell may be transformed with a vector comprising anucleic acid encoding the discrete AT. In addition, the host cell mayinclude a different vector comprising a nucleic acid encoding the ACPdomain.

[0030] In one embodiment, the extender molecule may be selected from thegroup consisting of malonyl CoA, alkyl malonyl CoA (e.g., methyl, ethyl,or propionyl malonyl CoAs), acyl inalonyl CoA, hydroxy malonyl CoA andalkoxy malonyl CoA (e.g., methoxy malonyl CoA). In another embodiment,the ACP domain may be selected from the group consisting of lnmI-ACP3,lmnJ-ACP4, lnmJ-ACP5, lnmJ-ACP6-1, lnmJ-ACP6-2, lnmJ-ACP7 and lnmJ-ACP8,descriptions of which are provided in a later Example. Furthermore, theACP domain may be present within the modular structure of a type I PKSor type I PKS/NRPS.

[0031] In another embodiment, the discrete AT provided in transcomprises a polypeptide selected from the group consisting of: acatalytic domain encoded by SEQ ID NO:43; a catalytic domain encoded bya nucleic acid having the sequence of a nucleic acid amplified by PCRusing the primer pair set forth in SEQ ID NO:155 and 156, respectively,using the nucleic acid of a leinamycin-producing organism as template;and a catalytic domain encoded by a nucleic acid that specificallyhybridizes to SEQ ID NO:43 under stringent conditions.

[0032] In yet another method according to the invention, the ACP domainis present within a type I PKS module lacking a naturally-occurring acyltransferase domain or includes an acyl transferase domain which has beenunnaturally rendered ineffective for loading the extender molecule tosaid ACP domain. The AT domain may also be selected from the group ofpolypeptides consisting of PksC/PksD/PksE-AT (from Bacillus subtilis),PedC/PedD (from a bacterial symbiont of Paederus beetles),MmpIII-AT1/AT2 (from Pseudomonas fluorescens), FenF (from Bacillussubtilis), Mx-TaK (from Myxococcus xanthus), LmnG (from Streptomycesatroolivaceus) or another Lnm-G homolog.

[0033] The present invention is also directed to methods of loading anextender molecule on a type I polyketide synthase (PKS) module,including the step of contacting said type I PKS module with theextender molecule and a recombinantly expressed discrete acyltransferase (AT), whereby the extender molecule is loaded onto the typeI PKS module by the acyl transferase catalytic activity of the discreteAT, provided in trans.

[0034] In yet another embodiment of the invention, methods for producinga polyketide or polyketide/nonribosomal peptide hybrid or analog areprovided. These methods include the steps of: providing a type Ipolyketide synthase (PKS) or type I PKS/nonribosomal protein synthetase(PKS/NRPS) including a module lacking an acyltransferase (AT) domain orincluding an acyltransferase domain which has been unnaturally renderedineffective for loading the extender molecule on said module; contactingthe PKS or NRPS with a recombinantly-expressed, discrete acyltransferase(AT) wherein the discrete AT is catalytically active in loading anextender molecule onto the module lacking in acyl transferase activity;incubating under conditions permitting the biosynthesis of thepolyketide or polyketide/nonribosomal peptide or analog thereof; andisolating the polyketide or polyketide/nonribosomal peptide or analogthereof. Such methods are preferably carried out in a host cell. Morepreferably, the host cell includes a vector comprising a nucleic acidencoding the discrete AT. Alternatively, the host cell includes anadditional but different vector comprising a nucleic acid encoding theACP domain.

[0035] Definitions

[0036] The term “polyketide synthases” (PKSs) refers to multifunctionalenzymes, related to fatty acid synthases (FASs). PKSs catalyze thebiosynthesis of polyketides through repeated (decarboxylative) Claisencondensations between acylthioesters, usually acetyl, propionyl, malonylor methylmalonyl. Following each condensation, they typically introducestructural variability into the product by catalyzing all, part, or noneof a reductive cycle comprising a ketoreduction, dehydration, andenoylreduction on the β-keto group of the growing polyketide chain. PKSsincorporate enormous structural diversity into their products, inaddition to varying the condensation cycle, by controlling the overallchain length, choice of primer and extender units and, particularly inthe case of aromatic polyketides, regiospecific cyclizations of thenascent polyketide chain. After the carbon chain has grown to a lengthcharacteristic of each specific product, it is typically released fromthe synthase by thiolysis or acyltransfer. Thus, PKSs consist offamilies of enzymes which work together to produce a given polyketide.Two general classes of PKSs exist. One class, known as Type I PKSs, isrepresented by the PKSs for macrolides such as erythromycin. These“complex” or “modular” PKSs include assemblies of several largemultifunctional proteins carrying, between them, a set of separateactive sites for non-iteratively carrying out each step of carbon chainassembly and modification (Cortes et al. (1990) Nature 348: 176; Donadioet al. (1991) Science 252: 675; MacNeil et al. (1992) Gene 115: 119).Structural diversity occurs in this class from variations in the numberand type of active sites in the PKSs. This class of PKSs displays aone-to-one correlation between the number and clustering of active sitesin the primary sequence of the PKS and the structure of the polyketidebackbone. The second class of PKSs, called Type II PKSs, is representedby the synthases for aromatic compounds. Type II PKSs typically have asingle set of iteratively used active sites (Bibb et al. (1989) EMBO J.8: 2727; Sherman et al. (1989 (EMBO J. 8: 2717; Fernandez-Moreno, et al.(1992) J. Biol. Chem. 267:19278).

[0037] A “nonribosomal peptide synthase” (NRPS) refers to an enzymaticcomplex of eukaryotic or prokaryotic origin, that is responsible for thesynthesis of peptides by a nonribosomal mechanism, often known asthiotemplate synthesis (Kleinkauf and von Doehren (1987) Ann. Rev.Microbiol., 41: 259-289). Such peptides, which can be up to 20 or moreamino acids in length, can have a linear, cyclic (cyclosporin,tyrocidine, mycobacilline, surfactin and others) or branched cyclicstructure (polymyxin, bacitracin and others) and often contain aminoacids not present in proteins or modified amino acids throughmethylation or epimerization.

[0038] A “module” refers to a set of distinctive polypeptide domainsthat encode all the enzyme activities necessary for one cycle ofpolyketide or peptide chain elongation and associated modifications.

[0039] The terms “isolated” “purified” or “biologically pure” refer tomaterial which is substantially or essentially free from componentswhich normally accompany it as found in its native state. With respectto nucleic acids and/or polypeptides the term can refer to nucleic acidsor polypeptides that are no longer flanked by the sequences typicallyflanking them in nature.

[0040] The terms “polypeptide”, “peptide” and “protein” are usedinterchangeably herein to refer to a polymer of amino acid residues. Theterms apply to amino acid polymers in which one or more amino acidresidue is an artificial chemical analogue of a corresponding naturallyoccurring amino acid, as well as to naturally occurring amino acidpolymers. The term also includes variants on the traditional peptidelinkage joining the amino acids making up the polypeptide.

[0041] The terms “nucleic acid” or “oligonucleotide” or grammaticalequivalents herein refer to at least two nucleotides covalently linkedtogether. A nucleic acid of the present invention is preferablysingle-stranded or double stranded and will generally containphosphodiester bonds, although in some cases, as outlined below, nucleicacid analogs are included that may have alternate backbones, comprising,for example, phosphoramide (Beaucage et al. (1993) Tetrahedron49(10):1925) and references therein; Letsinger (1970) J. Org. Chem.35:3800; Sprinzl et al. (1977) Eur. J. Biochem. 81: 579; Letsinger etal. (1986) Nucl. Acids Res. 14: 3487; Sawai et al. (1984) Chem. Lett.805, Letsinger et al. (1988) J. Am. Chem. Soc. 110: 4470; and Pauwels etal. (1986) Chemica Scripta 26: 1419), phosphorothioate (Mag et al.(1991) Nucleic Acids Res. 19:1437; and U.S. Pat. No. 5,644,048),phosphorodithioate (Briu et al. (1989) J. Am. Chem. Soc. 111 :2321,O-methylphophoroamidite linkages (see Eckstein, Oligonucleotides andAnalogues: A Practical Approach, Oxford University Press), and peptidenucleic acid backbones and linkages (see Egholm (1992) J. Am. Chem. Soc.114:1895; Meier et al. (1992) Chem. Int. Ed. Engl. 31: 1008; Nielsen(1993) Nature, 365: 566; Carlsson et al. (1996) Nature 380: 207). Otheranalog nucleic acids include those with positive backbones (Denpcy etal. (1995) Proc. Natl. Acad. Sci. USA 92: 6097; non-ionic backbones(U.S. Pat. Nos. 5,386,023, 5,637,684, 5,602,240, 5,216,141 and4,469,863; Angew. (1991) Chem. Intl. Ed. English 30: 423; Letsinger etal. (1988) J. Am. Chem. Soc. 110:4470; Letsinger et al. (1994)Nucleoside & Nucleotide 13:1597; Chapters 2 and 3, ASC Symposium Series580, “Carbohydrate Modifications in Antisense Research”, Ed. Y. S.Sanghui and P. Dan Cook; Mesmaeker et al. (1994), Bioorganic & MedicinalChem. Lett. 4: 395; Jeffs et al. (1994) J. Biomolecular NMR 34:17;Tetrahedron Lett. 37;743 (1996)) and non-ribose backbones, includingthose described in U.S. Pat. Nos. 5,235,033 and 5,034,506, and Chapters6 and 7, ASC Symposium Series 580, Carbohydrate Modifications inAntisense Research, Ed. Y. S. Sanghui and P. Dan Cook. Nucleic acidscontaining one or more carbocyclic sugars are also included within thedefinitions of nucleic acids (see Jenkins et al. (1995), Chem. Soc. Rev.pp169-176). Several nucleic acid analogs are described in Rawls, C & ENews Jun. 2, 1997 page 35. These modifications of the ribose-phosphatebackbone may be done to facilitate the addition of additional moietiessuch as labels, or to increase the stability and half-life of suchmolecules in physiological environments.

[0042] The term “heterologous” as it relates to nucleic acid sequencessuch as coding sequences and control sequences, denotes sequences thatare not normally associated with a region of a recombinant construct,and/or are not normally associated with a particular cell. Thus, a“heterologous” region of a nucleic acid construct is an identifiablesegment of nucleic acid within or attached to another nucleic acidmolecule that is not found in association with the other molecule innature. For example, a heterologous region of a construct could includea coding sequence flanked by sequences not found in association with thecoding sequence in nature. Another example of a heterologous codingsequence is a construct where the coding sequence itself is not found innature (e.g., synthetic sequences having codons different from thenative gene). Similarly, a host cell transformed with a construct whichis not normally present in the host cell would be consideredheterologous for purposes of this invention.

[0043] A “coding sequence” or a sequence that “encodes” a particularpolypeptide (e.g. a PKS, an NRPS, etc.), is a nucleic acid sequencewhich is ultimately transcribed and/or translated into that polypeptidein vitro and/or in vivo when placed under the control of appropriateregulatory sequences. In certain embodiments, the boundaries of thecoding sequence are determined by a start codon at the 5′ (amino)terminus and a translation stop codon at the 3′ (carboxy) terminus. Acoding sequence can include, but is not limited to, cDNA fromprokaryotic or eukaryotic mRNA, genomic DNA sequences from prokaryoticor eukaryotic DNA, and even synthetic DNA sequences. In preferredembodiments, a transcription termination sequence will usually belocated 3′ to the coding sequence.

[0044] Expression “control sequences” refers collectively to promotersequences, ribosome binding sites, polyadenylation signals,transcription termination sequences, upstream regulatory domains,enhancers, and the like, which collectively provide for thetranscription and translation of a coding sequence in a host cell. Notall of these control sequences need always be present in a recombinantvector so long as the desired gene is capable of being transcribed andtranslated.

[0045] “Recombination” refers to the reassortment of sections of DNA orRNA sequences between two DNA or RNA molecules. “Homologousrecombination” occurs between two DNA molecules which hybridize byvirtue of homologous or complementary nucleotide sequences present ineach DNA molecule.

[0046] The terms “stringent conditions” or “hybridization understringent conditions” refers to conditions under which a probe willhybridize preferentially to its target subsequence, and to a lesserextent to, or not at all to, other sequences. “Stringent hybridization”and “stringent hybridization wash conditions” in the context of nucleicacid hybridization experiments such as Southern and northernhybridizations are sequence dependent, and are different under differentenvironmental parameters. An extensive guide to the hybridization ofnucleic acids is found in Tijssen (1993) Laboratory Techniques inBiochemistry and Molecular Biology—Hybridization with Nucleic AcidProbes part I chapter 2 Overview of principles of hybridization and thestrategy of nucleic acid probe assays, Elsevier, New York. Generally,highly stringent hybridization and wash conditions are selected to beabout 5° C. lower than the thermal melting point (T_(m)) for thespecific sequence at a defined ionic strength and pH. The T_(m) is thetemperature (under defined ionic strength and pH) at which 50% of thetarget sequence hybridizes to a perfectly matched probe. Very stringentconditions are selected to be equal to the T_(m) for a particular probe.

[0047] An example of stringent hybridization conditions forhybridization of complementary nucleic acids which have more than 100complementary residues on a filter in a Southern or northern blot is 50%formamide with 1 mg of heparin at 42 C., with the hybridization beingcarried out overnight. An example of highly stringent wash conditions is0.15 M NaCl at 72 C. for about 15 minutes. An example of stringent washconditions is a 0.2×SSC wash at 65 C. for 15 minutes (see, Sambrook etal. (1989) Molecular Cloning—A Laboratory Manual (2^(nd) ed.) Vol. 1-3Cold Spring Harbor Laboratory, Cold Spring Harbor Press, NY, for adescription of SSC buffer). Often, a high stringency wash is preceded bya low stringency wash to remove background probe signal. An examplemedium stringency wash for a duplex of, e.g., more than 100 nucleotides,is 1×SSC at 45 C. for 15 minutes. An example low stringency wash for aduplex of, e.g., more than 100 nucleotides, is 4-6×SSC at 40 C. for 15minutes. In general, a signal to noise ratio of 2× (or higher) than thatobserved for an unrelated probe in the particular hybridization assayindicates detection of a specific hybridization. Nucleic acids which donot hybridize to each other under stringent conditions are stillsubstantially identical if the polypeptides which they encode aresubstantially identical. This occurs, e.g., when a copy of a nucleicacid is created using the maximum codon degeneracy permitted by thegenetic code.

[0048] A “library” or “combinatorial library” of polyketides and/orpolypeptides is intended to mean a collection of polyketides and/orpolypeptides (or other molecules) catalytically produced by a PKS and/orNRPS and/or hybrid PKS/NRPS (or other possible combination of syntheticelements) gene cluster. The library can be produced by a gene clusterthat contains any combination of native, homolog or mutant genes fromaromatic, modular or fungal PKSs and/or NRPSs. The combination of genescan be derived from a single PKS and/or NRPS gene cluster, e.g., act,fren, gra, tcm, whiE, gris, ery, or the like, and may optionally includegenes encoding tailoring enzymes which are capable of catalyzing thefurther modification of a polypeptide, polyketide, or other molecule.Alternatively, the combination of genes can be rationally orstochastically derived from an assortment of NRPS and/or PKS geneclusters. The library of polyketides and/or polypeptides and/or othermolecules thus produced can be tested or screened for biological,pharmacological or other activity.

[0049] By “random assortment” is intended any combination and/or orderof genes, homologs or mutants which encode for the various PKS and/orNRPS enzymes, modules, active sites or portions thereof derived fromaromatic, modular or fungal PKS and/or NRPS gene clusters.

[0050] By “genetically engineered host cell” is meant a host cell wherethe native PKS and/or NRPS gene cluster has been altered or deletedusing recombinant DNA techniques or a host cell into which heterologousPKS and/or NRPS and/or hybrid PKS/NRPS gene cluster has been inserted.Thus, the term would not encompass mutational events occurring innature. A “host cell” is a cell derived from a prokaryotic microorganismor a eukaryotic cell line cultured as a unicellular entity, which canbe, or has been, used as a recipient for recombinant vectors bearing thePKS, NRPS, and/or hybrid gene clusters of the invention. The termincludes the progeny of the original cell which has been transfected. Itis understood that the progeny of a single parental cell may notnecessarily be completely identical in morphology or in genomic or totalDNA complement to the original parent, due to accidental or deliberatemutation. Progeny of the parental cell which are sufficiently similar tothe parent to be characterized by the relevant property, such as thepresence of a nucleotide sequence encoding a desired PKS, are includedin the definition, and are covered by the above terms.

[0051] “Expression vectors” are defined herein as nucleic acid sequencesthat direct the transcription of cloned copies of genes/cDNAs and/or thetranslation of their mRNAs in an appropriate host. Such vectors can beused to express genes or cDNAs in a variety of hosts such as bacteria,bluegreen algae, plant cells, insect cells and animal cells. Expressionvectors include, but are not limited to, cloning vectors, modifiedcloning vectors, specifically designed plasmids or viruses. Specificallydesigned vectors allow the shuttling of DNA between hosts, such asbacteria-yeast or bacteria-animal cells. An appropriately constructedexpression vector preferably contains: an origin of replication forautonomous replication in a host cell, as electable marker, optionallyone or more restriction enzyme sites, optionally one or moreconstitutive or inducible promoters. In preferred embodiments, anexpression vector is a replicable DNA construct in which a DNA sequenceencoding a one or more PKS and/or NRPS domains and/or modules isoperably linked to suitable control sequences capable of effecting theexpression of the products of these synthase and/or synthetases in asuitable host. Control sequences include a transcriptional promoter, anoptional operator sequence to control transcription and sequences thatcontrol the termination of transcription and translation, and so forth.

[0052] A “leinamycin (Lnm) open reading frame”, or “ORF”, or “Lnm Orf”refers to a nucleic acid open reading frame that encodes a polypeptideor polypeptide domain that has an enzymatic activity used in thebiosynthesis of a leinamycin.

[0053] A “PKS/NRPS/PKS” system refers to a synthetic system comprisingan NRPS flanked by two PKSs. A “NRPS/PKS/NRPS” system refers to asynthetic system comprising a PKS flanked by two NRPSs. A “hybridPKS/NRPS system” or a “hybrid NRPS/PKS system” refers to a hybridsynthetic system comprising at least one PKS and one NRPS module. Thesystem can comprise multiple modules and the order can vary.

[0054] A “biological molecule that is a substrate for a polypeptideencoded by a leinamycin biosynthesis gene” refers to a molecule that ischemically modified by one or more polypeptides encoded by open readingframe(s) of the Lnm gene cluster. The “substrate” may be a nativemolecule that typically participates in the biosynthesis of aleinamycin, or can be any other molecule that can be similarly actedupon by the polypeptide.

[0055] A “polymorphism” is a variation in the DNA sequence of somemembers of a species. A polymorphism is thus said to be “allelic,” inthat, due to the existence of the polymorphism, some members of aspecies may have the unmutated sequence (i.e. the original “allele”)whereas other members may have a mutated sequence (i.e. the variant ormutant “allele”). In the simplest case, only one mutated sequence mayexist, and the polymorphism is said to be diallelic. In the case ofdiallelic diploid organisms, three genotypes are possible. They can behomozygous for one allele, homozygous for the other allel orheterozygous. In the case of diallelic haploid organisms, they can haveone allele or the other, thus only two genotypes are possible. Theoccurrence of alternative mutations can give rise to trialleleic, etc.polymorphisms. An allele may be referred to by the nucleotide(s) thatcomprise the mutation.

[0056] “Single nucleotide polymorphism” or “SNPs” are defined by theircharacteristic attributes. A central attribute of such a polymorphism isthat it contains a polymorphic site, “X,” most preferably occupied by asingle nucleotide, which is the site of the polymorphism's variation(Goelet and Knapp U.S. patent application Ser. No. 08/145,145). Methodsof identifying SNPs are well known to those of skill in the art (see,e.g., U.S. Pat. No. 5,952,174).

[0057] “Homologous”, as used herein, refers to the sequence similaritybetween two polypeptide molecules or between two nucleic acid molecules.When a position in both of the two compared sequences is occupied by thesame base or amino acid monomer subunit, e.g., if a position in each oftwo DNA molecules is occupied by adenine, then the molecules arehomologous at that position. The percent of homology between twosequences is a function of the number of matching or homologouspositions shared by the two sequences divided by the number of positionscompared times 100. For example, if 6 of 10, of the positions in twosequences are matched or homologous then the two sequences are 60%homologous. By way of example, the DNA sequences ATTGCC and TATGGC share50% homology. Generally, a comparison is made when two sequences arealigned to give maximum homology.

[0058] The following abbreviations are used herein: A, adenylation; ACP,acyl carrier protein; AT, acyltransferase, lnm, leinamycin; C,condensation; CL, co-enzyme A ligase; Cy, condensation/cyclization; DH,dehydratase; ER, enoyl reductase; KR, ketoreductase; KS, ketoacylsynthase; MT, methyltransferase; NRPS, nonribosomal peptide synthetase;orf, open reading frame; Ox, oxidation; PCP, peptidyl carrier protein;PCR, polymerase chain reaction; PKS, polyketide synthase; ACP, arylcarrier protein, bp, base pair, CoA, co-enzyme A, DTT, dithiothreitol;FAS, fatty acid synthase; kb, kilobase; PPTase, 4′-phosophopantetheinyltransferase; TCA, triloroacetic acid; TE, thioesterase.

BRIEF DESCRIPTION OF THE DRAWINGS

[0059]FIG. 1 illustrates the structure of leinamycin (Lnm) and theproposed mechanism for its oxidative and alkylative DNA-cleavage.

[0060]FIG. 2 shows a map of the 132 kb DNA from S. altroolivaceus thatharbors the leinamycin biosynthetic gene cluster.

[0061]FIG. 3 shows the genetic organization n of the leinamycinbiosynthetic gene cluster from S. altroolivaceus. NRPS and PKS genes areshown by solid arrows.

[0062]FIG. 4 illustrates a proposed Lnm biosynthetic pathway in S.altroolivaceus involving a hybrid NRPS/PKS. Abbreviations: A,adenylation; ACP, acyl carrier protein; AT, acyltransferase; C,condensation; CL, co-enzyme A ligase; Cy, condensation; cyclization; DH,dehydratase; ER, enoyl reductase; KR, ketoreductase; KS, ketoacylsynthase; Ox, oxidation; MT, methyltransferase; PCP, peptidyl carrierprotein; TE, thioesterase.

[0063]FIG. 5 alignment of Streptomyces lnm pptase with pptase fragmentsfrom other species. SC5A7 (SEQ ID NO:223), color (SEQ ID NO:224), Svp(SEQ ID NO:225), gris (SEQ ID NO:226), albua (SEQ ID NO:227), lnm (SEQID NO:228), consensus (SEQ ID NO:229).

[0064]FIG. 6 illustrates how altering lnmG expression alters leinamycinexpression. Inactivation of lnmG yields an S. atroolivaceus mutantstrain whose ability to produce leinamycin is completely abolished.Introduction of an lnmG overexpression plasmid into the S. atroolivaceuslnmG mutant not only restores its ability to produce leinamycin but alsoresults in an overproduction of leinamycin in comparison with the wildtype S. atroolivaceus strain. Thus, S. atroolivaceus lnmG mutanttransformed with a low-copy-number (10) plasmid in which the expressionof lnmG is under the control of the ermE* promoter produces similarlevel of leinamycin as the wild type S. atroolivaceus strain. S.atroolivaceus lnmG mutant transformed with a medium-copy-number (300)plasmid in which the expression of lnmG is under the control of theermE* promoter produces 3-5 fold more leinamycin than the wild type S.alroolivaceus strain.

[0065]FIG. 7 illustrates examples of novel Lnm analogs that can beprepared by manipulating the Lnm NRPS and PKS genes.

[0066]FIG. 8 illustrates nucleophilic attack of the episulfonium ion by—NH₂ or H₂O via S_(N)1 or S_(N)2 mechanism.

[0067] FIGS. 9-9C illustrate the production of novel leinamycins byengineering leinamycin biosynthesis. FIG. 9A: LnmH is a protein ofunknown function on the basis of amino acid sequence analysis.Inactivation of LnmH yields an S. alroolivaceus mutant that no longerproduces leinamycin but accumulates at least two new leinamycinmetabolites upon HPLC analysis. Complementation of the LnmH mutant byoverexpression of LnmH under the ermE* promoter in a low-copy-numberplasmid restores the leinamycin production to the mutant strain with thesame metabolite profile as the wild type S. atroolivaceus strain. InFIG. 9B, leinamycin productions were compared in fermentation mediasupplemented with various concentrations of D-alanine. Leinamycinproduction can be improved by 3-5 folds upon addition of 25 mMD-analine. FIG. 9C showed that inactivation of either one of the twoP-450 hydroxylases (LnmA or LnmZ) can lead to the production of newleinamycins, one of which could be the 8-dehydroxyl-Lnm showed in FIG.7.

[0068]FIG. 10 shows the results of targeted gene inactivation of lnmGand lnmI on leinamycin biosynthesis and establishes the involvement ofthe cloned gene cluster in leinamycin production. LnmG is a di-domainproteins that shows high amino acid sequence homology to knownpolyketide synthase. LnmI is multi-domain protein that shows high aminoacid sequence homology to known polyketide synthase and nonribosomalpeptide synthetase. In activation of lnmG and lnmI, respectively, bytargeted gene replacement experiments, produces S. atroolivaceus mutantstrains that no longer produce leinamycin and its biosyntheticintermediates. This was confirmed by HPLC analysis.

[0069]FIG. 11 shows LNM (1) biosynthesis hypothesis and modularorganization of the Lnm hybrid NRPS-PKS megasynthetase with a discreteLnmG AT enzyme loading the malonyl CoA extender to all six PKS modules.Structures in brackets are hypothetic. It is not known if one or bothACPs in module-6 are loaded with the malonyl group in vivo, althoughLnmG prefers ACP6-2 in vitro. Abbreviations are: A, adenylation; ACP,acyl carrier protein; AT, acyltransferase; Cy, condensation/cyclization;DH, dehydratase; KR, ketoreductase; KS, β-ketoacyl synthase; MT, methyltransferase; NRPS, nonribosomal peptide synthetase; Ox, oxidation; PCP,peptidyle carrier protein; PKS, polyketide synthase; TE, thioesterase.

[0070]FIG. 12 illustrates HPLC analysis of LNM production by wild-typeand recombinant S. atroolivaceus strains. (A) LNM standard, (B) S-140,(C) SB3001 (ΔlnmI), (D) SB3002 (ΔlnmJ), (E) SB3003 (ΔlnmG), (F) SB3004(SB3003 harboring the lnmG overexpression plasmid of pBS3017), (G)SB3005 (SB3003 harboring the lnmG overexpression plasmid of pBS3018).(♦), LNM; and (∇), an unknown metabolite whose production is independentto LNM biosynthesis.

[0071]FIG. 13 depicts in vitro assays of LnmG-catalyzed loading ofmalonyl CoA to individual LnmIJ PKS ACP domains and the LnmJ-(DH-ACP-KR)tridomain protein. (A) Purified LnmG on 4-15% sodium dodecylsulfate-polyacrylaminde gel (SDS-PAGE). Lane 1, molecular weightstandards (MW Stds); lane 2, LnmG. (B) Purified LnmIJ ACPs and LnmP on4-15% SDS-PAGE. Lane 1, MW Stds; lane 2, ACP3; lane 3 ACP4; lane 4,ACP5; lane 5, ACP6-1; lane 6, ACP6-2; lane 7, ACP7; lane 8, ACP8; lane9, LnmP. The numbers after the ACPs refer to PKS modules from which theyare derived with 6-1 and 6-2 to indicate the first and second ACPs,respectively, for PKS module 6. (C) Incubation of holo-ACPs or PCP with[2-¹⁴C]malonyl CoA and LnmG as visualized on 4-15% SDS-PAGE (I) and byphosphor imaging (II). Lane 1, MW Stds; lanes 2 to 8; ACP3 to ACP8; lane9, LnmP. (D) Time course of LnmG-catalyzed loading of [2-¹⁴C]malonyl CoAto ACP3 as visualized on 4-15% SDS-PAGE (I) and by phosphor imaging(II). (E) HPLC analysis of LnmG-catalyzed loading of malonyl CoA toACP3. I, a negative control in the absence of Svp; II, a negativecontrol in the absence of LnmG; III, complete assay. (•), apo-ACP3; (∇),holo-ACP3; (♦), malonyl-S-ACP3. F. Purified LnmJ-(DH-ACP-KR) on 9%SDS-PAGE. Lane 1, MW Stds, lane 2, LnmJ-(DH-ACP-KR). G. Incubation ofholo-LnmJ-(DH-ACP-KR) with [2-¹⁴C]malony CoA and LnmG as visualized on9% SDS-PAGE (I) and by phosphor imaging (II). Lane 1, MW Stds, lane 2, anegative control in the absence of Svp; lanes 3-6, complete assay withincubation time of 2, 5, 15, and 60 min, respectively.

[0072]FIG. 14 depicts a phylogenetic analysis of discrete ATs andcognate AT domains of type I polyketide synthases and type I polyketidesynthase/nonribosomal peptide synthetases performed by the CLUSTAL Wprogram (version 1.81), using Gonnet series weight matrix with a gapopen penalty of 10.00 and a gap extension penalty of 0.2. (Higgins, D.G., Thompson, J. D. and Gibson, T. J. (1996); Methods Enzymol., 266,383-402).

[0073]FIG. 15 depicts multiple sequence alignment performed by theCLUSTAL W program (version 1.81), using Gonnet series weight matrix witha gap open penalty of 10.00 and a gap extension penalty of 0.2.

DETAILED DESCRIPTION

[0074] This invention pertains to the isolation, identification, andcharacterization of the gene clusters that directs synthesis ofLeinamycin (Lnm). In addition, the invention pertains to uses of variouselements of the Lnm cluster, in particular, LnmG. Leinamycin is amacrolactam of hybrid polyketide and nonribosomal peptide origin with anunprecedented 1,3-dioxo-1,2-dithiolane structure. Lnm shows potentantitumor activity in tumor models in vivo, including those that areresistant to clinically important anticancer drugs. Although Lnm analogswith improved antitumor activity have been generated by chemicalmodifications, supporting the wisdom of making novel anticancer drugsbased on the Lnm scaffold, development of Lnm into a clinical anticancerdrug has been hampered by the in vivo instability of natural Lnm.

[0075] Genetic manipulations of both polyketide and nonribosomal peptidebiosynthesis have generally been very successful in generating novel“unnatural” natural products. In a similar manner, extension of thesegenetic-based approaches, also known as “Combinatorial Biosynthesis,” toLnm biosynthesis provides the ability to generate novel Lnm analogs,which are expected to provide useful “lead compounds” for thedevelopment of novel anticancer drugs.

[0076] The leinamycin synthetic pathway described herein utilizes ahybrid polyketide-peptide biosynthesis. Polyketides and polypeptides canbe assembled in a remarkably similar manner by repetitive addition of anextending unit to a growing chain by polyketide synthases (PKS) andnonribosomal peptide synthetase (NRPS) respectively. In the case ofpolyketides, the extending unit is typically a fatty acid (activated asan acyl CoA thioester) while the extending unit for polypeptides istypically an amino acid (activated as an aminoacyl adenylate). Both thePKS and NPRS systems have evolved a modular organization to define thenumber, sequence, and specificity of the incorporation of the extendingunit and utilized the 4′-phosphopanththeine prosthetic group to channelthe growing intermediate during the elongation process.

[0077] Polyketide metabolites display enormous structural diversity, yetshare a common mechanism of biosynthesis. The carbon backbones ofpolyketides are assembled from short carboxylic acids by sequentialdecarboxylative condensation, and this process is catalyzed by PKSs. Twotypes of bacterial PKSs have been characterized to explain thepolyketide biochemistry (Katz and Donadio (1993) Ann. Rev. Microbiol.47: 875-912; Hutchinson and Fuji, (1995) Ann. Rev. Microbiol. 49:201-38; Cane (1997) Chem. Rev. 97: 2463-2705; Carreras et al. (1997)Top. Curr. Chem. 188: 85-126; Cane, et al. (1998) Science 282: 63-68;Staunton and Wilkinson (1998) Top. Curr. Chem. 195:49-92; Hopwood andSherman (1990) Ann. Rev. Genet. 24: 37-66). Type I enzymes aremultifunctional proteins that harbor sets of noniteratively useddistinct active sites, termed modules, for the catalysis of each cycleof polyketide chain elongation in biosynthesis of reduced polyketideslike macrolide, polyether, or polyene antibiotics (Cane (1997) Chem.Rev. 97: 2463-2705; Carreras et al. (1997) Top. Curr. Chem. 188: 85-126;Staunton and Wilkinson (1998) Top. Curr. Chem. 195: 49-92). Type IIenzymes are multienzyme complexes that carry a single set ofiteratively-used activities and consist of several monofunctionalproteins for the synthesis of aromatic polyketides like tetracyclines(Cane (1997) Chem. Rev. 97: 2463-2705). The growing polyketideintermediates in both systems remain covalently attached to the acylcarrier protein (ACP) of the PKS enzyme via the 4′-phosphopantetheinecofactor during the elongation process (Shen, et al. (1997) J.Bacteriol., 174: 3818-3821; Carreras and Khosla (1998) Biochemistry 37:2084-2088).

[0078] Nonribosomal peptides, a structurally diverse family of bioactivepeptides, are assembled nonribosomally from both proteinogenic andnonproteinogenic amino acids, and this process is catalyzed by NRPSs.Remarkably, NRPS possess a similar multimodular structure as type IPKSs, and these modules represent the functional building units of anNRPS that activate, modify and link together by amide or ester bonds theconstituent amino acids of the peptide product (Cane (1997) Chem. Rev.97: 2463-2705; Carreras et al. (1997) Top. Curr. Chem. 188: 85-126;Cane, et al. (1998) Science 282: 63-68; Cane and Walsh (1999) Chem.Biol. 6: R319-R325; Konz and Marahiel (1999) Chem. Biol. 6: R39-R48; vonDöhren et al. (1999) Chem. Biol. 6: R273-R279). Modules can be eitherphysically linked together, or interact noncovalently viaprotein/protein recognition to form the protein template that dictatesthe number and sequence of the amino acids incorporated into the peptideproducts. Individual modules are characterized by domains that catalyzethe activation of constituent amino acids as acyladenylates (A domain)(Weinreb et at. (1998) Biochemistry 37:1575-1584: Stachlhaus andMarahiel (1995) J. Biol. Chem. 270: 6163-6169; Konz et al. (197) Chem.Biol. 4: 927-937; Mootz and Marahiel (1997) J. Bacteriol. 179:6843-6850; Stachelhaus et al. (1999) Chem. Biol., 6: 493-505; Challis etal. (2000) Chem. Biol. 7: 211-224), the thioesterification of theactivated amino acids with the sulthydryl group of the4′-phosphopantetheine cofactor that is covalently bound to theC-terminal region of each amino acid activating domain (thiolationdomain or PCP) (Gehring et al. (1997) Chem. Biol. 4: 17-24; Lambalot etal. (1996) Chem. Biol. 3:923-936; Weinreb et al. (1998) Biochemistry 37:1575-1584; Stachlhaus and Marahiel (1995) J. Biol. Chem. 270: 6163-6169;Konz et al. (197) Chem. Biol. 4:927-937; Mootz and Marahiel (1997) J.Bacteriol. 179: 6843-6850; Stachelhaus et al. (1999) Chem. Biol., 6:493-505; Challis et al. (2000) Chem. Biol. 7:211-224; Stachelhaus et al.(1996) Chem. Biol. 3: 913-921; Pfeifer et al. (1995) Biochemistry, 34:7450-7459; Haese et al. (1994) J. Mol. Biol. 243: 116-122; Quadri et al.(1998) Bilchemistry, 37: 1585-1595; Rose et al. (1998) Nucleic AcidsRes., 26: 1628-1635; Ku et al. (1997) Chem. Biol., 4: 203-207; Reuter etal. (1999) EMBO J., 18: 6823-6831), and the transpeptidation of carboxythioester activated amino acids between the aligned domains, resultingin the formation of a specific peptide chain with a defined sequence(condensation or C domain) (Stachihaus et al. (1998) J. Biol. Chem.,273: 22773-22781).

[0079] Additional domains have also been identified that are responsiblefor the modification of the peptide product, such as epimerization (E)domains for the conversion of L-amino acids to D-amino acids (Marahielet al. (1997) Chem. Rev., 97: 2651-2673), N-methyltransferase (MT)domains for the addition of methyl groups to the amide nitrogen (Id.),cyclization (Cy) domains for the formation of heterocyclic rings (Konzet al. (197) Chem. Biol. 4: 927-937), a reduction domain for reductiverelease of an aldehyde product (Ehmann et al. (1999) Biochemistry, 38:6171-6177), and oxidation domains for the thiazolidine-to-thiazoleoxidation (Ox) (Molnar et al. (1999) Chem. Biol., 7: 97-109) or forα-hydroxylation of the incorporated amino acid (Ox′) (Silakowski et al.(1999) J. Biol. Chem., 274: 37391-37399).

[0080] Hybrid peptide-polyketide metabolites, such as leinamycin, arestructurally characterized by both the amino acid and the carboxylicacid building blocks. The assembly of hybrid peptide-polyketidemetabolites from amino acids and carboxylic acids is catalyzed by ahybrid NRPS-PKS system that bears the characteristics of both NRPS andPKS (Du et al. (2001) Metabolic Eng. 3: 78-95; Du and Shen (2001) Curr.Opinion Drug Discov. Dev. 4: 215-228). The interacting NRPS and PKSmodules can be either covalently linked by arranging the catalyticdomains in a linear order on the same protein, or physically located ontwo or more separate proteins, utilizing specific protein-proteinrecognition to ensure the correct pairing between the interactingmodules. One such hybrid system is exemplified by the bleomycinbiosynthesis, BlmIX NRPS/Blm VIII PKS/BlmVII NRPS system, combining thefeatures of both hybrid NRPS/PKS and PKS/NRPS systems (see e.g. USSNPCT/US00/0045; Shen et al. (1999) Bioorg. Chem., 27: 155-171).

[0081] In one aspect, this invention provides a cloned and characterized(LNM) leinamycin gene cluster (˜61.3 kb) consisting of characteristicNRPS and PKS genes from the (lnm) leinamycin producer Streptomycesatroolivaceus. The cloned and isolated (lnm) leinamycin gene cluster andsubunits (e.g. ORFs) thereof provides a method of recombinantlyexpressing leinamycin and/or leinamycin analogues. Thus, in oneembodiment, this invention provides for nucleic acids encodingleinamycin synthetic machinery or subunits thereof, for cellsrecombinantly modified to express a leinamycin and/or leinamycinanalogue, and for a leinamycin or leinamycin analogue recombinantlyexpressed in such cells.

[0082] Like other polyketide synthase or nonribosomal peptidesynthetases, the leinamycin synthetic pathway is organized into modules,each module catalyzing the addition and/or modification of one subunit(e.g. fatty acid and/or amino acid). Each module is organized into anumber of domains each domain having a characteristic activity (e.g.activation, condensation, condensation, cyclization, etc.). Thecatalytic domain within a module and the modules themselves are oftenarranged collinearly and the order of biosynthetic modules from NH₂- toCOOH-terminus on each PKS and NRPS polypeptide and the number and typeof catalytic domains within each determine the order of structural andfunctional elements in the resulting product.

[0083] The size and complexity of the ultimately formed product arecontrolled, in part, by the number of repeated acyl chain extensionsteps that are, in turn, a function of the number and placement ofcarrier protein domains in these multimodular enzymes. The numbercomposition and order of such domains can be altered either to introducemodifications, e.g. into the leinamycin to produce leinamycin analogues,or to produce different or completely new molecules. Such“recombination” is not restricted solely to recombination among theleinamycin catalytic domains and/or modules, but can also involverecombination between leinamycin modules and/or subunits and other PKSand/or NRPS modules and/or subunits (e.g. bleomycin subunits). Moreoverthe discovery that synthetic pathways can incorporate both PKS and NRPSmodules and/or catalytic domains makes available hybrid PKS/NRPSsynthases.

[0084] Thus, in one embodiment this invention contemplates the use oflnm gene cluster modules and/or catalytic domains to make variouspeptide and/or polyketide, and/or hybrid polypeptide/polyketidemetabolites (including, but not limited to leinamycin, leinamycinintermediates or shunt metabolites), in combinatorial biosynthesis withother polyketide synthases and/or other nonribosomal peptidesynthetases.

[0085] In particular, it is noted that the various lnm ORFs showcharacteristic enzymatic activities including, but not limited toaminotransferases, peptide synthetases, thioesterases, decarboxylases,and the like. The proteins encoded by these orfs can be used alone, orin combination with other active domains to modify various targetsubstrates.

[0086] This invention also includes the discovery and characterizationof a novel PPTase (a fragment of which is shown and named lmn in FIG.5). This PPTase can be cloned and used in engineered biosynthesis ofpolyketides, peptides, hybrid peptide and polyketide metabolites, hybridpolyketide and peptide metabolites, or the combination of both types ofmetabolites. The PPTase can also be used in converting apo-peptidylcarrier proteins (both type I and type II) and acyl carrier proteins(both type I and type II) into the holo-proteins.

[0087] In certain preferred embodiments, this invention contemplates theuse of leinamycin (lnm) gene cluster modules and/or catalytic domains toproduce leinamycin (e.g to upregulate endogenous leinamycin productionto permit leinamycin production in cells other than Streptomyces, etc.)and/or to make veious modified leinamycins (leinamycin analogues).

[0088] The Examples provided herein and the accompanying primers permitone of ordinary skill in the art to isolate the (lnm) leinamycin genecluster of this invention, its constituent orfs, various modules, orenzymatic domains. The isolated nucleic acid components can be used toexpress one or more polypeptide components for in vivo (e.g.recombinant) synthesis of one or more polypeptides and/or polyketides asindicated above. It will also be appreciated that the (lnm) leinamycincluster polypeptides can be used for ex vivo assembly of variousmacromolecules.

[0089] I. Biosynthetic Pathway for Leinamycin in S. atroolivaceus.

[0090] Without being bound to a particular theory, it is believed thatthe genetic organization of the lnm gene cluster and the enzymology ofLnm biosynthesis closely parallels that of macrolide and hybridpeptide-polyketide metabolites such as bleomycin. The structuralsimilarity of Lnm to macrolides in general and to thiazole-containinghybrid peptide-polyketide metabolites such as bleomycin in particularprovides substantial data on a pathway for Lnm biosynthesis. We believethe synthesis of the Lnm inacrolactam backbone involves a hybridNRPS/PKS system consisting of 2 NRPS and 6 PKS modules, which, at eachstep, specify the amino acid or carboxylic acid to be incorporated, andthe modifications associated with this particular cycle of elongation(FIG. 4). In such a hybrid NRPS/PKS biosynthesis (FIG. 4), Lnmbiosynthesis starts with an NRPS module, NRPS-1, that activatesL-alanine as an amino acylthioester and epimerizes the latter intoD-alanine to set the stage for Lnm biosynthesis. In certain embodiments,however, D-alanine can be a direct substrate for NRPS-1. NRPS-2generally specifies L-cysteine is typically characterized by thesignature Cy and Ox domains for thiazole formation. The NRPS-2-boundpeptidyl intermediate is next elongated by the PKS-1 module, the KSdomain of which is homologous to the KSs known to interact with NPRSmodules in NRPS/PKS hybrids (PCT/US00/00445) and the ACP domain of whichis loaded by the acyltransferace (AT) domain of lnmG that should specifyfor malonyl CoA as an extender unit (Haydock et al. (1995) FEBS Lett.,374: 246-248).

[0091] Chain elongation proceeds by sequential incorporation of fiveadditional molecules of malonyl CoA, utilizing five additional PKSmodules, PKS-2, PKS-3, PKS-4, PKS-5, and PKS-6. The loading of malonylCoA to the ACP domains of all five PKS modules is catalyzed by the sameAT domain of lnmG whose substrate specificity for malonyl CoA as theextender unit for individual PKS modules is easily determined fromsequence analysis of the AT domains (Haydock et al. (1995) FEBS Lett.,374: 246-248). Since the AT domain of lnmG calls for malonyl CoA as anextender unit for all six PKS modules, an additional hydroxylation stepmust have taken place for the introduction of the methyl group at C-6.Macrolide ring hydroxylation is often catalyzed by cytochrome P-450hydroxylases that have been identified from several macrolidebiosynthesis gene clusters (Rodriguez et al. (1995) FEMS Microbiol.Lett., 127: 117-120; Weber et al. (1991) Science, 252: 114-117; Xue etal. (1998) Chem. Biol,. 5: 661-667).

[0092] Candidates for these types of hydroxylases have indeed beenidentified for leinamycin biosynthesis, such as lnmA, lnmB, and lnmZ(FIG. 4). Methylation of macrolide ring has been noted before, derivingthe methyl group from S-adenosylmenthionine (AdoMet) and catalyzed by amethyl transferase domain (Due et al (2000) Chem. Biol., 7: 623-642). AnMT domain has been identified in PKS4 of lnmJ, which presumably isresponsible for the introduction of the C-6 methyl group of leinamycin(FIG. 4). Finally, modeled on rafamycin biosynthesis—the onlymacrolactam whose biosynthesis gene cluster has been characterized todate (August et al. (1998) Chem. Biol., 5:69-79), we propose that adiscrete amide synthase catalyzes the release of the full lengthpeptide/polyketide intermediate from the Lnm synthase complex and thecyclization of the linear intermediate into macrolactam.

[0093] II. Leinamycin (lnm) gene cluster.

[0094] The nucleic acids comprising the leinamycin (lnm) gene clusterare identified in Table 2 and listed in the sequence listing providedherein (SEQ ID NO:1). In particular, Table 1 identifies genes andfunctions of open reading frames (ORFs) responsible for the biosynthesisof leinamycin, while Table 2 identifies a number of ORFs comprising theleinamycin (lnm) gene cluster, identifies the activity of the catalyticdomain encoded by the ORF and provides primers for the amplification andisolation of that orf.

[0095] As illustrated in Example 1, the leinamycin (lnm) clustercomprises NRPS modules followed by several PKS modules as well asseveral resistance and regulatory genes (Table 1). TABLE 1 Summary offunction of open reading frames (ORFs) in the leinamycin biosyntheticgene cluster and flanking regions. Sequence ID Nos refer to amino acidsequences encoded by ORF. # SED Amino Sequence homologue ID ORF acidsProposed function (Genbank accession no) NO −35 289 Probably antibioticresistance protein SpcN (AAD50455) 2 −34 502 Putative FAD-dependentoxygenase EncM (AAF81732) 3 −33 1237 Subtilisin-like secreted proteaseSAM-P45 (BAA12040) 4 −32 262 Unknown 5 −31 401 Probably NADHdehydrogenase II (E75456) 6 −30 306 RNA polymerase ECF-type sigma factorBH0672 (BAB04391) 7 −29 327 Probable macrolide 2'-phosphotransferaseMphA (D16251) 8 −28 198 Probable tetR-family transcriptional regulator(T37015) 9 −27 538 Antibiotic efflux protein ActVA-1 (S18539) 10 −26 300Putative hydroxylase SnoaW (AAF01810) 11 −25 197 Probable cyanamidehydratase (P22143) 12 −24 353 Histidinol-phosphate aminotransferase(P45358) 13 −23 774 Putative transcriptional regulator (T34847) 14 −2272 MbtH-like protein (AAG02368) 15 −21 1105 Nonribosomal peptidesynthetase (T30289) 16 −20 330 Probable regulatory protein SyrP (U88574)17 −19 335 Probable regulatory protein SyrP (U88574) 18 −18 313 Unknown19 −17 341 Putative fatty acid desaturase (AAA99932) 20 −16 433Diaminopimelate (DAP) decarboxylase (P00861) 21 −15 794 Putativepeptidase (NP422131) 22 −14 432 Antibiotic transport protein SpcT(AAD50454) 23 −13 1134 Nonribosomal peptide synthetase (T30289) 24 −12276 Conserved, function known (NP_421851) 25 −11 549 Nonribosomalpeptide synthetase NosA (AF204805) 26 −10 235 Thioesterase (AAC01736) 27−9  276 Short-chain dehydrogenase/reductase LimC (CAB54559) 28 −8  920Nonribosomal peptide sythetase AcmB (T14591) 29 −7  195 ProbableN-carbamoyl-sarcosine amidase Ta0454 (CAC11596) 30 −6  343 Hydrogenaseexpression/formation protein HypE (P24193) 31 −5  791 Hydrogenasematuration protein (regulator) HypF (P30131) 32 −4  447 Serinehydroxmethyl-transferase (SHMT) GlyA (029406) 33 −3  238 Probableglutamine amidotransferase (C83609) 34 −2  1745 Nonribosomal peptidesynthetase BlmVI (AF210249) 35 −1  462 Nonribosomal peptide synthetasePvdD (S53999) 36 lnmA 399 Cytochrome P450 hydroxylase RapN (T30231) 37lnmB 78 Ferredoxin (T30230) 38 lnmC 115 Unknown 39 lnmD 438 Probable3-oxoadipate enol-lactone PcaL (AAC38246) 40hydrolase/4-carboxymuconolactone decarboxylase lnmE 307 Unknown 41 lnmF265 Probable enoyl-CoA hydratase PksH (P40805) 42 lnmG 795 Probablemalonyl-CoA acyltransferase/enoyl FenF (T44805) 43 reductase lnmH 274Unknown 44 lnmI 4437 Hybrid nonribosomal peptide MtaC/MtaB (AF188187) 45synthetase/polyketide synthase lnmJ 7349 Polyketide synthase/ MtaB(AF188287)/ 46 Tyrosine phenol-lyase/ketoadipate-enol lactone PcaL(AAC38246) hydrolase lnmK 319 Conserved, function known TaD (CAB46503)47 lnmL 86 Acyl carrier protein (ACP, type II) TaE (CAB46504) 48 InmM416 ACP synthase (FabH homolog) TaF (CAB46505) 49 lnmN 267 Thioesterase(type II) GrsT (P14686) 50 lnmO 227 Probable transcriptional activatorNtcA (AAC14592) 51 lnmP 82 Peptidyl carrier protein (PCP, type II)(CAB99152) 52 lnmQ 516 Nonribosomal peptide synthetase (A-domain(AAG02343) 53 only, type II) lnmR 575 ABC transporter component (ATPhydrolase) MoaD (T45539) 54 lnmS 287 ABC transporter component (membraneAgaC (T45530) 55 spanning protein) lnmT 321 ABC transporter component(membrane AgaB (T45531) 56 spanning protein) lnmU 513 ABC transportercomponent (periplasmic OphA (S77572) 57 oligopeptide binding protein)lnmV 120 Unknown 58 lnmW 516 4-coumarate:CoA ligase (T08074) 59 lnmX 243Conserved, function known (CAC04222) 60 lnmY 474 Antibiotic effluxprotein Mct (AAD32747) 61 lnmZ 400 Cytochrome P450 hydroxylase MycG(SS1594) 62 lnmZ′ 134 Unknown 63 +1 216 Conserved, function known(C70555) 64 +2 272 Thioesterase GrsT(P14686) 65 +3 345 Conserved,function known (CAC18692) 66 +4 236 Probable tetR-family transcriptionalregulator HemR (BAA21913) 67 +5 539 Antibiotic resistance protein CarA(AAC32027) 68 +6 322 Probable hydrolase/lactonase VgbB (AAC61670) 69 +7551 ABC transporter VarM (BAA96297) 70 +8 469 AdenosylhomocysteinaseSahH (CAB88907) 71 +9 303 5,10-methylenetetrahydrofolate reducatase MetF(O54253) 72

[0096] TABLE 2 ORFs, deduced functions, amino acid sequence homologs,and PCR primers for amplification of individual ORFs identified in theleinamycin biosynthetic gene cluster and its flanking regions SEQProtein ID ORF# Position Proposed Function Homologue Primers NO −353489- Probable antibiotic resistance SpcN Fwd: ATGGAGATGTCCGACACC 734358 protein (AAD50455) Rev CTACTGGCCGCTGCCCAG 74 −34 5108- PutativeFAD-dependent EncM Fwd: ATGAGCGACTTTTCCCGC 75 6616 oxygenase (AAF81732)Rev: TCAGCGGGACGCAGGCGG 76 −33 7431- Subtilisin-like secreted SAM-P45Fwd: TTGCCCAAGCTTCCCATC 77 11144 protease (BAA 12040) Rev:TCAGCGCAGGCCGAAGGC 78 −32 11141- Unknown Fwd: ATGGCGGACGAACCTGCG 7911812 Rev: TCATCGTTCCGTCCTCCT 80 −31 11809- Probable NADH (E75456) Fwd:ATGAGCGCACGGCAGGAG 81 13014 dehydrogenase II Rev: TCACCGTGCCTCCCGGAC 82−30 13011- RNA polymerase ECF- type BH0672 Fwd: GTGACCGACCCGACCGCC 8313931 sigma factor (BAB04391) Rev: TCAGCGCGTCCCGACGTC 84 −29 14271-Probable macrolide 2′- MphA Fwd: ATTGGTTGCGAACGAGGGT 85 15254pbospbotransferase (D16251) Rev: TCAGCCGAAGCGGCGGAA 86 −28 16277-Probable tetR-family (T37015) Fwd: ATGGGCCGCGTGTCCCAG 87 15681transcriptional regulator Rev: TCAGTCCATGCGCTGCTG 88 −27 16467-Antibiotic efflux ActVA-1 Fwd: GTGGCATCGCCACCCACC 89 18083 protein(S18539) Rev: TCACTTGTCACCGCCGGT 90 −26 18480- Putative hydroxylaseSnoaW Fwd: ATGACTGCCGACAACCTG 91 19382 (AAF01810) Rev:TCAGCCCAGGTAGAGGTC 92 −25 20377- Probable cyanamide (P22143) Fwd:ATGACACTGGACCACCTG 93 19784 hydratase Rev: TCAGTCGAGGCTGTTGGT 94 −2420662- Histidinol-phosphate (P45358) Fwd: TTGACCACGCTCACGTTC 95 21723aminotransferase Rev: TCACCGCACGAACGCGTT 96 −23 21994- Putativetranscriptional (T34847) Fwd: ATCCAATTCAGCTCGCGA 97 24318 regulator Rev:TCAGGAGCCCGCGGCCAC 98 −22 24524- MbtH-like protein (AAG02368) Fwd:ATGAGCGATCGGGACAGT 99 24742 Rev: TCAGCTGCGGCCCGCCTG 100 −21 24778-Nonribosomal peptide (T30289) Fwd: ATGCAGACCCAGCTCTCC 101 28095synthetase Rev: TCAGCGGCGCTGCGCGCC 102 −20 28128- Probable regulatorySyrP Fwd: ATGACCATTGAGGTGCAC 103 29120 protein (U88574) Rev:TCATGCGGGCACCTCGCC 104 −19 29117- Probable regulatory protein SyrP Fwd:ATGACGCTCACCGACCTG 105 30124 (U88574) Rev: TCATCGGCCGGCCGGCAG 106 −1830172- Unknown Fwd: ATGCTGCTGCGCCCCACC 107 31113 Rev: TCAGCCGGCCGGGGCCGA108 −17 31140- Putative fatty acid desaturase (AAA99932) Fwd:ATGACGCAGACCGCCCCC 109 32165 Rev: TCACGTCCACGGCGTGCT 110 −16 32199-Diaminopimelate (DAP) (P00861) Fwd: ATGAGACCCGACATGAGT 111 33500Decarboxylase Rev: TCACAGACCCTCGGGGAT 112 −15 35984- Putative peptidase(NP-422131) Fwd: ATGGCCGACACCCGTACC 113 33600 Rev: TCAGAGCACGTATCGGCG114 −14 37313- Antibiotic transport protein SpcT Fwd: GTGGCGCCGCGCACGCCG115 36015 (AAD50454) Rev: TCAGGTCCGTTCCGGTGC 116 −13 40721- Nonribosomalpeptide (T30289) Fwd: ATGACCGAGACCCTGCCC 117 37317 synthetase Rev:TCAGCCCTCCAGCTTCTG 118 −12 41548- Conserved, function (NP_421851) Fwd:ATGCGATCCGTCCGCACC 119 40718 unknown Rev: TCATCGCTGTCCCTCCGC 120 −1141709- Nonribosomal peptide NosA Fwd: ATGACGGCCGACGATTCG 121 43358synthetase (AF204805) Rev: TCAGGCGGGCGCCTGTTC 122 −10 43412-Thioesterase (AAC01736) Fwd: ATGTTGAGTGCGGCGGTT 123 44119 Rev:TCATGACGGCGTCCCGGC 124  −9 44116- Short-chain LimC Fwd:ATGAGCGGACGGCTCACG 125 44946 dehdrogenase/reductase (CAB54559) Rev:TCAACGGGCGCTGTAGCC 126  −8 44970- Nonribosomal peptide AcmB (T14591)Fwd: GTGTCGTCCAACTCCCCT 127 47732 synthetase Rev: TCAGGCCGTCCTCGCCGC 128 −7 47820- Probable N-carbamoyl- Ta0454 Fwd: ATGAGCAAGGTCGCGGTC 12948407 sarcosine amidase (CAC11596) Rev: TCAGGGGGTGCGGAACAC 130  −648545- Hydrogenase expression/ HypE Fwd: TTGCCGACGGCCACGACG 131 49576formation protein (P24193) Rev: CAGCACAGGCGGGGAAG 132  −5 49599-Hydrogenase maturation HypF Fwd: ATGGCAGAGACCGAGCAG 133 51974 protein(regulator) (P30131) Rev: TCAGCGGCATTCGTTCGT 134  −4 52006- Serinehydroxymethyl- GlyA Fwd: ATCCGGACCGCAGATCTG 135 53349 transferase (SHMT)(029406) Rev: TCACCGGGACGCCTCTGT 136  −3 53346- Probable glutamine(C83609) Fwd: GTGAGCCGGCCGGTCATC 137 54062 amidotransferase Rev:TCAGACGGATGCCGCTGT 138  −2 54157- Nonribosomal peptide BlmV1 Fwd:GTGCACACTCACGTCCGT 139 59394 synthetase (AF210249) Rev:TCAGCCTTGCTGCTGCAG 140  −1 59420- Nonribosomal peptide PvdD Fwd:ATGGCCGTGACACTCAAG 141 60808 synthetase (S53999) Rev: TCAACTCACCGCCGGCTG142 InmA 60948- Cytochrome P450 RapN Fwd: ATGTCGGCTACGAGGCGG 143 62147hydroxylase (T30231) Rev: TCACCATGCGATCGGCAG 144 InmB 62159- Ferredoxin(T30230) Fwd: ATGGCACGGGAGCAGAAC 145 62395 Rev: TCACGACAGGTCGAGCAC 146InmC 62682- Unknown Fwd: ATGAAGTTCGCGATCGTC 147 63029 Rev:TTACTCGGCCACCCACAG 148 InmD 63116- Probable 3-oxoadipate PcaL Fwd:ATGACGGACGGCGCGATA 149 64432 enol-lactone (AAC38246) Rev:TCACCGTGCGGCGCCGCT 150 hydrolase/4-carboxy- muconolactone decarboxylaseInmE 64500- Unknown Fwd: ATGACCGACGCGGCGAGC 151 65423 Rev:TCAGAACCAGGCGGGCGC 152 InmF 65441- Probableenoyl-CoA PksH Fwd:GTGACGGCCATCGGCCCG 153 66238 hydratase (P40805) Rev: TCAGGGCCGCGGCTTCTC154 InmG 66268- Probable malonyl-CoA FenF Fwd: ATGGTGGCACTGGTTTTC 15568655 acyltransferase/enoyl (144805) Rev: TCAGCGGCGGGCGAGGAC 156reductase InmH 68725- Unknown Fwd: ATGACCACCCTGACCTTC 572 69549 Rev:CTAGCGGGCGTCCGGCAC 158 InmI 69681- Hybrid nonribosomal peptide MtaC/MtaBFwd: ATGACCACCCTGACCTTC 159 82994 synthetase/ (AF188187) Rev:TCACCACTTCCGTCCTTC 160 polyketide synthase InmJ 82991- Hybrid polyketideMtaB Fwd: GTGAACGTGCCCTCCGCA 161 105040 synthase/tyrosine (AF188287)/Rev: TCATGCCGGGTGCTCCTC 162 phenol-lyase PeaL ketoadipate-enol(AAC38246) lactone hydrolase InmK 105037- Conserved, function TaD Fwd:ATGACCATCACCTCGTCG 163 105996 unknown (CAB46503) Rev: TCATGCTTCCCCCTTCGG164 InmL 105993- Acyl carrier protein TaE Fwd: ATGACCCAGGCACCACTG 165106253 (ACP, type II) (CAB46504) Rev: TCATCGCGGGGCTCCGCT 166 InmM106250- ACP synthase (FabH TaF Fws: ATGACCGCGACCGGTGCC 167 107500homolog) (CAB46505) Rev: TCAGCGCCACGCGTACTG 168 InmN 107557-Thioesterase (type II) GrsT Fwd: GTGTACGGCTCTCGGACG 169 108360 Rev:TCACGTGGCAACTTTATG 170 InmO 108395- Probable transcriptional NtcA Fwd:ATGAACCTGCTGGATGTC 171 109078 activator (AAC14592) Rev:TCAGACGCATCGGCTCTC 172 InmP 109122- Peptidyl carrier protein (PCP,(CAB99152) Fwd: ATGTGGGACCACAAGTTC 173 109370 type II) Rev:TCATCGGCCGGCTCCGTC 174 InmQ 109367- Nonribosomal peptide (AAG02343) Fwd:ATGAGCGGCGCCAAGCTG 175 110917 synthetase (A-domain) Rev:TCAGGACGCCGGGGCGAG 176 InmR 112700- ABC transporter component MoaD Fwd:TTGAGCGCAGTCTTCGAC 177 110973 (T45539) Rev: TCAGACCCCGTCGACTGC 178 InmS113560- ABC transporter component AgaC Fwd: ATGACGGCCCCGACGCCG 1791112697 (T45530) Rev: TCAAGGCACGAACCTCGC 80 InmT 114522- ABC transportercomponent AgaB Fwd: GTGACGTCCGCCGTCCGG 181 113557 (T45531) Rev:TCATGTCGCCGTCCTCAT 182 InmU 116060- ABC transporter component OphA Fwd:ATGTCACGGGTCAACGGC 183 114519 (S77572) Rev: TCACGCGGACCTGGCCCG 184 InmV116494- Unknown Fwd: ATGAGCACCGACAGGAG 185 116132 Rev.TCAGGCCCACCAGTCGCG 186 lnmW 118041- 4-coumarate: CoA (T08074) Fwd:ATGACGGAACGGACGTTC 187 116491 ligase Rev: TCATGACGGGGCTCCTGT 188 InmX118780- Conserved, function (CAC04222) Fwd: ATGGCCGACACACTCCTC 189118049 unknown Rev: TCAACCCACTATCTGGAA 190 InmY 120239- Antibioticefflux protein Mct Fwd: ATGACCGTCAGGACCGAC 191 118815 (AAD32747) Rev:TCAGGCGGCGGCGTCGGT 192 InmZ 121638- Cytochrome P450 MycG Fwd:GTGAGCACCGAAGTGGAA 193 120436 hydroxylase (S51594) Rev:TCACCACTCGACGTGCAT 194 InmZ 121757- Unknown Fwd: ATGACTCAGATGCGGATT 195122161 Rev: CTAGGCAGCCCCGTCGGT 196  +1 122832- Conserve, functionunknown (C70555) Fwd: ATGGCGCCCGGCTCCGGC 197 122182 Rev:TCAGCCCTTCCCGGCCGC 198  +2 123664- Thioesterase GrsT Fwd:GTGGACCGAGAGGGGAAC 199 122846 (P14686) Rev: TCAGAACGTCCGCTCGGC 200  +3123898- Conserved, function (CC18692) Fwd: ATGACCGGCACGCTCGTG 201 124935unknown Rev: TCACCAACTGGTCCTGCT 202  +4 125516- Probable tetR-familyHemR Fwd: ATGCCACCGCCTCCCCGA 203 124806 transcriptional regulator(BAA21913) Rev: TCAGATCAGGGCGCGCCG 204  +5 125637- Antibiotic resistanceprotein CarA Fwd: ATGCCTACGCAGATCAGC 205 127256 (AAC32027) Rev:TCAGACCCGGACGGCCTG 206  +6 127231- Probable VgbB Fwd: ATGCACCACAGGCCGTCC207 128199 hydrolase/lactonase (AAC61670) Rev: CTAGAGCTCCATGCGCAG 208 +7 129971- ABC transporter VarM Fwd: ATGACCACCCACCCGAAC 209 128316(BAA96297) Rev: TCACGGTGTCACCGCTTC 210  +8 131727- Adenosylhomocysteinase SahH Fwd: ATGCCCTCGCAGCCGCCC 211 130318 (CAB88907) Rev:TCAGTAGCGGTAGTGGTC 212  +9 132616- 5,10-methylenetetra- MetF Fwd:TTGAGCACGCTGCGCGAC 213 131705 hydrofolate reductase (054253) Rev:TCAGCGGGCGGCTGCGAG 214

[0097] This invention also provides the cloning and sequence of a4′-phophopantetheinyl transferase (PPTase) from S. atroolivaceus. ThePPTase sequence from S. alroolivaceus is named as “lnm” in the aminoacid pileup analysis illustrated in FIG. 5. The rest are PPTases fromother peptide and/or polyketide producing microorganisms. The Lnm PPTasedescribed herein facilitates engineered biosynthesis of the leinamycinNRPS-PKS genes for generation of chemical structural diversity.

[0098] Using the sequence information provided herein (e.g. primersequences and PPTase sequence) the PPTase nucleic acids can be routinelyisolated according to standard methods (e.g. PCR amplification).

[0099] III. Expression of Leinamycin (Inm) Gene Clusters, Modules, andEnzymatic Domains.

[0100] As indicated above, in one embodiment this invention providesnovel NRPS and PKS genes for the efficient recombinant production ofboth novel and known polyketides, peptides, and polyketide/polypeptidehybrids by expressing them in vivo. In other embodiments, such synthesesare carried out in vitro. Even in vitro syntheses, however, typicallyutilize recombinantly expressed PKSs,NRPSs, or enzymatic domainsthereof. Thus, it is frequently desirable to express protein componentsof the PKSs or NRPs described above.

[0101] Typically expression of the protein components of the pathwayand/or of the products of the NRPS/PKS pathway is accomplished byplacing the subject PKS or NRPS nucleic acid(s) in an expression vector,and transfecting a cell with the vector such that the cell expressed thedesired product(s).

[0102] A Isolation/Preparation of Nucleic Acids.

[0103] In one embodiment, this invention provides nucleic acids for therecombinant expression of a leinamycin. Such nucleic acids includeisolated gene cluster(s) comprising open reading frames encodingpolypeptides sufficient to direct the assembly of a leinamycin.

[0104] In other embodiments of this invention, modified leinamycins(e.g. leinamycin analogs), novel polyketides, polypeptides, andcombinations thereof (polyketide/polypeptide hybrids) are created bymodifying PKSs and/or NRPSs so as to introduce variations into knownpolymers synthesized by the enzymes. Such variation may be introduced bydesign, for example to modify a known molecule in a specific way, e.g.by replacing a single monomeric unit within a polymer with another,thereby creating a derivative molecule of predicted structure.Alternatively, variations can be made randomly, for example by making alibrary of molecular variants of a known polymer by systematically orhaphazardly replacing one or more modules or enzymatic domains in aknown PKS or NRPS with a collection of alternative modules or domains.Production of alternative/modified PKSs, NRPSs and hybrid systems isdescribed below.

[0105] Using the primer and sequence information provided herein, one ofordinary skill in the art can routinely isolate/clone the leinamycin PKSand/or NRPS modules and/or enzymatic domains (ORFs) described herein.For example, the PCR primers provided in Table 2, above, can be used toamplify any of the orfs identified therein. Moreover, using the sequenceinformation for the leinamycin (lnm) gene cluster provided herein (see,e.g., SEQ ID NO:1), the design of other primers suitable of theamplification of individual ORFs, combinations of ORFs, genes, etc. isroutine.

[0106] Typically such amplifications will utilize the DNA of an organismcontaining the requisite genes (e.g. Streptomyces atroolivaceus) as atemplate. Typical amplification conditions include a PCR mixtureconsisting of 5 ng of S. atroolivaceus genomic or plasmid DNA astemplate, 25 pmoles of each primers, 25 μM dNTP, 5% DMSO, 2 units of Taqpolymerase, 1× buffer, with or without 20% glycerol in a final volume of50 μL. PCR is carried out (e.g. on a Gene Amp PCR System 2400(Perkin-Elmer/ABI)) with a cycling scheme as follows: initial denaturingat 94° C. for 5 min, 24-36 cycles of 45 sec at 94° C., 1 min at 60° C.,2 min at 72° C., followed by additional 7 min at 72° C. One of skillwill appreciate that optimization of such a protocol, e.g. to improveyield, etc. is routine (see e.g., U.S. Pat. No. 4,683,202; Innis (1990)PCR Protocols A Guide to a Methods and Applications. Academic Press Inc.San Diego, Calif., etc.). In addition, primer may be designed tointroduce restriction sites and so facilitate cloning of the amplifiedsequence into a vector.

[0107] Using the information provided herein other approaches to cloningthe desired sequences will be apparent to those of skill in the art. Forexample, the PKS or NRPS modules or enzymatic domains of interest can beobtained from an organism that expresses the same, using recombinantmethods, such as by screening cDNA or genomic libraries, derived fromcells expressing the gene, or by deriving the gene from a vector knownto include the same. The gene can then be isolated and combined withother desired NRPS and/or PKS modules or domains, using standardtechniques. If the gene in question is already present in a suitableexpression vector, it can be combined in situ, with, e.g., other PKSand/or NRPS subunits, as desired. The gene of interest can also beproduced synthetically, rather than cloned. The nucleotide sequence canbe designed with the appropriate codons for the particular amino acidsequence desired. In general, one will select preferred codons for theintended host in which the sequence will be expressed. The completesequence can be assembled from overlapping oligonucleotides prepared bystandard methods and assembled into a complete coding sequence (see,e.g., Edge (1981) Nature 292:756; Nambair et al. (1984) Science 223:1299; Jay et al. (1984) J. Biol. Chem. 259:6311). In addition, it isnoted that custom gene synthesis is commercially available (see, e.g.Operon Technologies, Alameda, Calif.).

[0108] Examples of such techniques and instructions sufficient to directpersons of skill through many cloning exercises are found in Berger andKimmel (1989) Guide to Molecular Cloning Techniques, Methods inEnzymology 152 Academic Press, Inc., San Diego, Calif. (Berger);Sambrook et al. (1989) Molecular Cloning—A Laboratory Manual (2nd ed.)Vol. 1-3, Cold Spring Harbor Laboratory, Cold Spring Harbor Press, NY;Ausubel (19 1994) Current Protocols in Molecular Biology, CurrentProtocols, a joint venture between Greene Publishing Associates, Inc.and John Wiley & Sons, Inc., U.S. Pat. No. 5,017,478; and EuropeanPatent No. 0,246,864.

[0109] B) Expression Vectors and Host Cells.

[0110] A wide variety of expression vectors and host cells are suitablefor the synthesis of leinamycin or leinamycin analogues, or theexpression of polypeptides comprising the leinamycin biosyntheticpathway.

[0111] The choice of vector depends on the sequence(s) that are to beexpressed. Any transducible cloning vector can be used as a cloningvector for the nucleic acid constructs of this invention. However, wherelarge clusters are to be expressed, phagemids, cosmids, P1s, YACs, BACs,PACs, HACs or similar cloning vectors can be used for cloning thenucleotide sequences into the host cell. Phagemids, cosmids, and BACs,for example, are advantageous vectors due to the ability to insert andstably propagate therein larger fragments of DNA than in M13 phage andlambda phage, respectively. Phagemids which will find use in this methodgenerally include hybrids between plasmids and filamentous phage cloningvehicles. Cosmids which will find use in this method generally includelambda phage-based vectors into which cos sites have been inserted.Recipient pool cloning vectors can be any suitable plasmid. The cloningvectros into which pools of mutants are inserted may be identical or maybe constructed to harbor and express different genetic markers (see,e.g., Sambrook et al., supra). The utility of employing such vectorshaving different marker genes may be exploited to facilitate adetermination of successful transduction.

[0112] In certain embodiments of this invention, vectors are used tointroduce PKS, NRPS, or NRPS/PKS genes or gene clusters into host (e.g.Streptomyces) cells. Numerous vectors for use in particular host cellsare well known to those of skill in the art. For example described inMalpartida and Hopwood, (1984) Nature, 309:462-464; Kao et al., (1994),Science, 265: 509-512; and Hopwood et al., (1987) Methods Enzymol.,153:116-166 all describe vectors for use in various Streptomyces hosts.

[0113] In a certain embodiment, Streptomyces vectors are used thatinclude sequences that allow their introduction and maintenance in E.coli. Such Streptomyces/E. coli shuttle vectors have been described(see, for example, Vara et al., (1989) J. Bacteriol., 171:5872-5881;Guilfoile & Hutchinson (1991) Proc. Natl. Acad. Sci. USA, 88:8553-8557.)

[0114]S. atroolivaceus is sensitive to thiostrepton (Thi) and apramycin(Apr). Thus, in one preferred embodiment the pGM60 (Muth et al. (1989)Mol. Gen. Genet., 219: 341-348), vector carrying the Thi^(R) marker, andpKC1139 (Bierman et al. (1992) Gene, 116: 43-49) vector, carrying theApr^(R) marker, are particularly well suited for expression of lnmnucleic acids. Introduction of plasmid DNA into S. atroolivaceus byeither polyethyleneglycol (PEG)-mediated transformation of protoplasts(Hopwood et al. (1985) Genetic manipulation of Streptomyces: alaboratory manual., John Innes Foundation: Norwich, UK) or byconjugation from E. coli S17 (Bierman et al. (1992) Gene, 116: 43-49)was successful, demonstrating the feasibility of manipulating Lnmbiosynthesis in S. atroolivaceus in vivo.

[0115] The gene sequences, or fragments thereof, which collectivelyencode an lnm gene cluster, one or more ORFs, one or more lnm modules,or one or more lnm enzymatic domains of this invention, can be insertedinto expression vectors, using methods known to those of skill in theart. Preferred expression vectors will include control sequencesoperably linked to the desired NRPS and/or PKS coding sequence orfragment thereof. Suitable expression systems for use with the presentinvention include systems that function in eukaryotic and prokaryotichost cells. However, as explained above, prokaryotic systems arepreferred, and in particular, systems compatible with Streptomyces spp.are of particular interest. Control elements for use in such systemsinclude promoters, optionally containing operator sequences, andribosome binding sites. Particularly useful promoters include controlsequences derived from PKS and/or NRPS gene clusters, such as one ormore act promoters. However, other bacterial promoters, such as thosederived from sugar metabolizing enzymes, such as galactose, lactose(lac) and maltose, will also find use in the present constructs.Additional examples include promoter sequences derived from biosyntheticenzymes such as tryptophan (trp), the beta-lactamase (bla) promotersystem, bacteriophage lambda PL, and T5. In addition, syntheticpromoters, such as the tac promoter (U.S. Pat. No. 4,551,433), which donot occur in nature also function in bacterial host cells. InStreptomyces, numerous promoters have been described includingconstitutive promoters, such as ermE and tcmG (Shen and Hutchinson,(1994) J. Biol. Chem. 269: 30726-30733), as well as controllablepromoters such as actI and actIII (Pleper et al., (1995) Nature, 378:263-266; Pieper et al., (1995) J. Am. Chem. Soc., 117: 11373-11374; andWiesmann et al., (1995) Chem. & Biol. 2: 583-589).

[0116] Other regulatory sequences may also be desirable which allow forregulation of expression of the PKS replacement sequences relative tothe growth of the host cell. Regulatory sequences are known to those ofskill in the art, and examples include those which cause the expressionof a gene to be turned on or off in response to a chemical or physicalstimulus, including the presence of a regulatory compound. Other typesof regulatory elements may also be present in the vector, fore example,enhancer sequences.

[0117] Selectable markers can also be included in the recombinantexpression vectors. A variety of markers are know which are useful inselecting for transformed cell lines and generally comprise a gene whoseexpression confers a selectable phenotype on transformed cells when thecells are grown in an appropriate selective medium. Such markersinclude, for example, genes that confer antibiotic resistance orsensitivity to the plasmid. Alternatively, several polyketides arenaturally colored and this characteristic provides a built-in marker forselecting cells successfully transformed by the present constructs.

[0118] The various lnm PKS and/or NRPS clusters or subunits of interestcan be cloned into one or more recombinant vectors as individualcassettes, with separate control elements, or under the control of,e.g., a single promoter. The PKS and/or NRPS subunits can includeflanking restriction sites to allow for the easy deletion and insertionof other PKS subunits so that hybrid PKSs can be generated. The designof such unique restriction sites is known to those of skill in the artand can be accomplished using the techniques described above, such assite-directed mutagenesis and PCR.

[0119] Methods of cloning and expressing large nucleic acids such asgene clusters, including PKS- or NRPS-encoding gene clusters, in cellsincluding Streptomyces are well known to those of skill in the art (see,e.g., Stutzman-Engwall and Hutchinson (1989) Proc. Natl. Acad. Sci. USA,86: 3135-3139;,Motamedi and Hutchinson (1987) Proc. Natl. Acad. Sci.USA, 84: 4445-4449; Grim et al. (1994) Gene, 151: 1-10; Kao et al.(1994) Science, 265: 509-512; and Hopwood et al. (1987) Meth. Enzymol.,153: 116-166). In some examples, nucleic acid sequences of well over 100kb have been introduced into cells, including prokaryotic cells, usingvector-based methods (see, for example, Osoegawa et al., (1998)Genomics, 52: 1-8; Woon et al., (1998) Genomics, 50: 306-316; Huang etal., (1996) Nucl. Acids Res., 24: 4202-4209).

[0120] Host cells for the recombinant production of the leinamycin,leinamycin analogues, leinamycin shunt metabolites, lnm modules, orfs,or catalytic domains, and the like can be derived from any organism withthe capability of harboring a recombinant PKS, NRPS or PKS/NRPS genecluster. Thus, the host cells of the present invention can be derivedfrom either prokaryotic or eukarydtic organisms. However, preferred hostcells are those constructed from the actinomycetes, a class of mycelialbacteria that are abundant producers of a number of polyketides andpeptides. A particularly preferred genus for use with the present systemis Streptomyces. Thus, for example, S. verticillus S. ambofaciens, S.avermitilis, S. atroolivaceus, S. azureus, S. cinnamonensis, S.coelicolor, S. curacoi, S. erythraeus, S. fradiae, S. galilaeus, S.glaucescens, S. hygroscopicus, S. lividans, S. parvulus, S. peucetius,S. rimosus, S. roseofulvus, S. thermotolerans, S. violaceoruber, amongothers, will provide convenient host cells for the subject invention,with S. coelicolor being preferred (see, e.g., Hopwood, D. A. andSherman, D. H. Ann. Rev. Geneet. (1990) 24:37-66, O'Hagan, D. ThePolyketide Metabolites (Ellis Horwood Limited, 1991), for a descriptionof various polyketide-producing organisms and their natural products).

[0121] Two of the common problems associated with heterologous geneexpression in E. coli are (1) the formation of inclusion bodies, whichrequires additional steps of solubilization and refolding and oftenleads to inactive enzymes, and (2) the inadequate posttranslationalprocessing of the resulting enzymes. These problems can be circumventedby expressing genes of Streptomyces origin in Streptomyces as indicatedabove. In preferred embodiments, cloning is performed using E.coli-Streptomyces shuttle vectors (e.g., pWHM3, pWHM601, pANT1200,pANT1201, pGM60, pKC1139). Using shuttle vectors, most of thesubclonings can be easily carried out in E. coli. As indicated above,particularly preferred vectors include pGM60, pKC1139. If controlledexpression of the target gene is desired, pMR5 (McDaniel et al. (1993)Science, 262: 1546-1550) is a good choice since it carries the actI andactIII promoter pair, the transcription of which is under the control ofthe actII-ORF4 transcriptional regulator. This vector has been used forthe expression of various PKS (McDaniel et al. (1999) Proc. Natl. Acad.Sci., USA, 96: 1846-1851; Pieper et al. (1995) Nature, 378: 263-266)genes, including the Blm NRPS and PKS genes very recently by us, in S.coelicolor and S. lividans.

[0122] In addition, a new family of 4′-phosphopantetheine transferaseshas been recently identified, which catalyze the posttranslationalmodification by the covalent attachment of the 4′-phosphopantetheinemoiety of CoA to a conserved serine residue of either the PCP domain ofNRPS or the ACP domain of PKS (Gehring et al. (1997) Chem. Biol., 4:17-24; Lambalot et al. (1996) Chem. Biol., 3: 923-936; Walsh et al.(1997) Curr. Opinion Chem. Biol. 1: 309-315). It is now possible tooverproduce functional holo-NRPS or PKS either in vivo by co-expressionof the NRPS or PKS gene and a 4′-phosphopantetheine transferase gene (Duand Shen (1999) Chem. Biol., 6: 507-517; Cox et al. (1997) FEBS Lett.,405: 267-272; Ku et al. (1997) Chem. Biol., 4: 203-207) or in vitro byphosphopantetheinylation of an apo-ACP or apo-PCP with CoA in thepresence of a 4′-phosphopantetheine transferase (Du and Shen (1999)Chem. Biol., 6: 507-517; Cox et al. (1997) FEBS Lett., 405: 267-272).Both in vivo and in vitro methods have been established in ourlaboratory (Du and Shen (1999) Chem. Biol., 6: 507-517) and can beapplied to the production of functional Lnm NRPS and PKS enzymes, shouldthe need of posttranslational modification arise.

[0123] In certain embodiments this invention may make use of geneticallyengineered cells that either lack PKS and/or NRPS genes or have theirnaturally occurring PKS and/or NRPS genes substantially deleted. Thesehost cells can be transformed with recombinant vectors, encoding avariety of PKS and/or NRPS gene clusters, for the production of activepolyketides. The invention provides for the production of significantquantities of product, e.g. a leinamycin, at an appropriate stage of thegrowth cycle. The leinamycin, leinamycin analogues, or other polyketide,peptide, or hybrid polyketide/peptide metabolites so produced can beused as therapeutic agents, to treat a number of disorders, depending onthe type of metabolites in question.

[0124] The vectors and host cells described above can be used to expressvarious protein components of the polyketide and/or polypeptidesynthetic modules for subsequent isolation and/or to provide abiological synthesis of one or more desired biomolecules (e.g.leinamycin, leinamycin analogues, and the like). Where leinamycin,and/or leinamycin analogues, and/or one or more proteins of theleinamycin (lnm) cluster are expressed (e.g. overexpressed) forsubsequent isolation and/or characterization, the proteins are expressedin any prokaryotic or eukaryotic cell suitable for protein expression.In one preferred embodiment, the proteins are expressed in E. coli.Overexpression of leinamycin in E. coli is described in Example 1.

[0125] In certain embodiments, a eukaryotic host cell is preferred (e.g.where certain glycosylation patterns are desired). Suitable eukaryotichost cells are well known to those of skill in the art. Such eukaryoticcells include, but are not limited to yeast cells, insect cells, plantcells, fungal cells, and various mammalian cells (e.g. COS, CHO HeLacells lines and various myeloma cell lines)

[0126] C) Protein/Polyketide Recovery.

[0127] Polypeptide and/or polyketide recovery is accomplished accordingto standard methods well known to those of skill in the art. Thus, forexample where lnm cluster proteins are to be expressed and isolated, theproteins can be expressed with a convenient tag to facilitate isolation(e.g. a His₆) tag. Other standard protein purification techniques aresuitable and well known to those of skill in the art (see, e.g., Quadriet al. (1998) Biochemistry 37: 1585-1595; Nakano et al. (1992) Mol. Gen.Genet. 232: 313-321, etc.).

[0128] Similarly where components (e.g. modules and/or enzymaticdomains) of the leinamycin cluster are used to express variousbiomolecules (e.g. polyketides, polypeptides, etc.) the desired productand/or shunt metabolite(s) are isolated according to standard methodswell know to those of skill in the art (see, e.g., Carreras and Khosla(1998) Biochemistry, 37: 2084-2088; Deutscher (1990) Methods inEnzymology Volume 182: Guide to Protein Purification, M. Deutscher, ed.,and the like). Hara et al. (1989) J. Antibiol. 42: 1768-1774 disclosesand effective culture system that, with minor modifications was used toexpress leinamycin (see Example 1). Purification of expressed leinamycinis also described in Example 1.

[0129] D) Optimized Expression System

[0130] Four methods are typically used for introduction of plasmid DNAinto Streptomyces species: PEG-mediated protoplast transformation,electroporation, conjugation, and phage infection. Standard protocolsare available in the Streptomyces laboratory manual (Hopwood et al.(1985) Genetic manipulation of Streptomyces: a laboratory manual., JohnInnes Foundation: Norwich, UK), and several different transformationsystems have been described for various Streptomyces species (Liu andShen (2000) Antimicrob. Agents Chemother., 44; 382-392; Lichenstein etal. (1990) Gene, 88: 81-86; Zang et al. (1997) J. Ferment. Bioeng., 83:217-221; Matsushima and Baltz (1985) J. Bacteriol., 162: 180-185;Garcia-Dominguez et al. (1987) App. Environ. Microbiol., 53: 1376-1381;Aidoo et al. (1990) J. Gen. Microbiol., 136: 657-662; Illing et al.(1989) J. Gen. Microbiol., 135: 2289-2297).

[0131] Example 1, describes a transfection system optimized for S.atroolivaceus. A conjugation approach was pursued, with surprisingsuccess. Spores (1×10⁹) are heat-shocked or 20 min at 42° C. instead of10 min at 50° C. followed by incubation at 30° C. for up to 6 hours. Thegermination of spores can be monitored by microscopic checks every 30min from 4 hours after heat-shock. E. coli S17-1 (bearing the desiredconstruct) culture is freshly prepared and conjugation is conducted onmodified ISP-4 medium. After incubation, e.g. at 28° C. for 5 days,apparent positive ex-conjugates are identified. We calculatedconjugation/integration efficiency for a non-self-replicating constructpYC12 as approximately 1.8×10⁻⁸, and the conjugation efficiency forself-replicating construct was 5×10⁻⁷.

[0132] IV. Synthesis of Leinamycin and Leinamycin Analogues.

[0133] In one embodiment this invention provides methods of synthesizingleinamycins and recombinantly synthesized leinamycins. As indicatedabove, this is generally accomplished by providing an organism (e.g. abacterial cell) containing sufficient components of the leinamycin genecluster to direct synthesis of a complete leinamycin and/or leinamycinanalogue.

[0134] In one embodiment, the entire leinamycin cluster, or a fragmentthereof (e.g. designed to introduce a modification into the clusterthrough homologous recombination) is cloned into a Streptomyces strain(e.g., S. lividans, S. atroolivaceus, or S. coelicolor). Kao etal.(1994) Science, 265: 509-512, have cloned the 30 kb DEBS genes fromSacc. erythmea into S. coelicolor and produced 6-deoxyerythronolide B inS. coelicolor and these methods can be used construct an expressionplasmid for heterologous expression of the leinamycin cluster. Thismethod involves the transfer of DNA between a temperature-sensitiveplasmid and a shuttle vector by means of a homologous doublerecombination event in E. coli (Sssio et al., (2000) Nature Biotechnol.18: 343-345).

[0135] In one preferred embodiment, the two ends spanning the leinamycincluster or fragment thereof, or recombinant construct, are cloned into atemperature-sensitive plasmid that is chloramphenicol resistant (CM^(R))such as pCK6. Streptomyces DNA is then rescued from a donor into thetemperature-sensitive recipient by co-transforming E. coil with theCm^(R) recipient plasmid and the apramycin resistant (Ap^(R)) pKC505donor cosmid that contains the desired construct, followed bychloramphenicol and apramycin selection at 30° C. Colonies harboringboth plasmids (Cm^(R), Ap^(R)) will be shifted to 44° C. onchloramphenicol and apramycin plates and only those cointegrates formedby a single recombination event between the two plasmids are viable.Surviving colonies are then propagated at 30° C. on Cm^(R) plates toselect for recombinant plasmids formed by the resolution of cointegratesthrough a second recombinant event. The desired construct is cloned intothe Cm^(R) temperature-sensitive plasmid and is ready to be moved intoany expression plasmid by a similar means of homologous recombinantevent.

[0136] Another system illustrated in Example 1, exploits the fact thatS. atroolivaceus grows very well at 30° C.; and doesn't grow attemperature beyond 35° C. In addition, it is highly sensitive to bothapromycin (Am) and thiostrepton (Thio). Thus, E. coli—Streptomycesshuttle vectors pOJ260 (suicide vector, Am^(R)) and pKC1139(self-replicating vector, Am^(R)) can be used to make the desired lmp orlmp-modification constructs.

[0137] The methods and constructs of this invention can be used to alterexpression of endogenous leinamycin. Using the lnm gene clusterinformation provided herein, one of skill in the art may regulate thesynthesis of endogenous leinamycin. In particular, the expression ofvarious ORFs comprising the lnm gene cluster may be increased ordecreased to alter leinamycin synthesis levels.

[0138] Methods of altering the expression of endogenous genes are wellknown to those of skill in the art. Typically such methods involvealtering or replacing all or a portion o the regulatory sequencescontrolling expression of the particular gene that is to be regulated.In a preferred embodiment, the regulatory sequences (e.g., the nativepromoter) upstream of one or more of the lnm ORFs are altered.

[0139] This is typically accomplished by the use of homologousrecombination to introduce a heterologous nucleic acid into the nativeregulatory sequences. To downregulate expression of one or more lnmORFs, simple mutations that either alter the reading frame or disruptthe promoter are suitable. To upregulate expression of the lnm ORF(s)the native promoter(s) can be substituted with heterologous promoter(s)that induce higher than normal levels of transcription. In aparticularly preferred embodiment, nucleic acid sequences comprising thestructural gene in question or upstream sequences are utilized fortargeting heterologous recombination constructs. The use of homologousrecombination to alter expression of endogenous genes is described indetail in U.S. Pat. No. 5,272,071, WO 91/09955, WO 93/09222, WO96/29411, WO 95/31560, and WO 91/12650.

[0140] In addition, or alternatively, constructs can be introduced thatexpress particular ORF at higher levels than in the wildtype organism.For example, leinamycin production yield improvement by engineeringleinamycin biosynthesis has been demonstrated using lnmG as an example.LnmG is a di-domain protein with amino acid sequence homology toacyltransferase (AT) the 1st domain) and enoyl reductase (ER) (the 2nddomain). Inactivation of lnmG yields an S. atroolivaceus mutant strainwhose ability to produce leinamycin is completely abolished. This resultunambiguously establishes that lnmG, and thereby the cloned genecluster, encodes leinamycin production.

[0141] Introduction of an lnmG overexpression plasmid into the S.atroolivaceus lnmG mutant not only restores its ability to produceleinamycin but also result in an overproduction of leinamycin incomparison with the S. atroolivaceus strain. This as shown in FIG. 6, S.atroolivaceus lnmG mutant transformed with a low-copy number (10)plasmid in which the expression of lnmG is under the control of the ermEpromoter produces similar level of leinamycin as the wild type S.atroolivaceus strain. S. atroolivaceus lnmG mutant transformed with amedium-copy-number (300) plasmid in which the expression of lnmG isunder the control of the ermE promoter produces 3-5 fold more leinamycinthan the wild type S. atroolivaceus strain.

[0142] In one certain embodiment, this invention provides methods ofsynthesizing modified leinamycins or leinamycin analogs. Typically, insuch embodiments, the leinamycin analogs are synthesized either byintroducing specific perturbations into individual NRPS and/or PKSenzymatic domains or modules, or by reprogramming the linear order inwhich the NRPS or PKS enzymatic domains and/or modules appear in theleinamycin gene cluster. The former will lead to leinamycin analogs withtargeted modifications at the leinamycin backbone and the latter willallow incorporation of other extension units in variable sequence intothe biosynthesis of leinamycin.

[0143] In preferred embodiments modification o the lnm gene cluster toyield leinamycin analogues is accomplished by one of two differentapproaches. In one approach, the lnm enzymatic domains and/or modulesare altered in a directed manner (i.e. they are changed in a preselectedway), while in another approach, random/haphazard alterations areintroduced into the lnm cluster and the resulting products are screenedto identify those with desired properties.

[0144] A) Synthesis of Leinamycin Analogs by Specific Engineering of thelnm Genes.

[0145] The lnm genes can be re-engineered by means of specific mutationsor by reprogramming the linear order of the NRPS or PKS enzymaticdomains or modules. In this approach, a wild-type lnm allele (ORF) isreplaced (ore recombined) with a mutant construct containing various lnmORFs in a different order. These mutants are introduced into andexpressed in an appropriate host (e.g., Streptomyces or in aheterologous host). Since both NRPSs (Stachelhaus et al. (1995) Science,269: 69-72) and PKSs (Donadio et al. (1993) Proc. Natl. Acad. Sci. USA,90: 7119-7123, Donadio et al. (1995) J. Am., Chem, Soc., 117: 9105-9106,Cortes et al. (1995) Science, 268: 1487-1489) have shown considerabletolerance to reprogramming, it is expected that these modifications ofthe lnm cluster will result in the production of leinamycin analogs withpredicted structural alterations.

[0146] Using this approach, rational manipulations of genes governingLnm biosynthesis allow preparation of novel Lnm analogs that areextremely difficult to prepare by chemical modifications or that presenta formidable task by total synthesis. Examples of such analogs include8-dehydroxyl-Lnm, 6-demethyl-Lnm, and 8-dehydroxyl-6-demethyl-Lnm (FIG.7), which can be generated inactivating the genes encoding the C-8hydroxylases, such as lnmA, lnmB, or lnmZ, and the MT domain of PKS-4 oflnmJ encoding the C-6 methyl transferase individually or both,respectively (FIG. 4, FIG. 7).

[0147] The stability of Lnm under aqueous condition depends on the pH:t_(1/2)>20 hr at pH 6, t_(1/2)=8 hr at pH 7, and t_(1/2)<1 hr at pH 8(Asai et al. (1997) Bioorg. Med. Chem. 5: 723-729). In the presence of athiol, Lnm exhibits an even shorter t_(1/2) and is inactivated bydegradation to form two major adducts A and B (FIG. 8). Removal of the8-hydroxyl group as in 8-dehydroxyl-Lnm should therefore eliminate theformation of adduct B, effectively enhancing the concentration of theactive form of episulfonium ion. Adduct A results from nucleophilicattack of the episulfonium ion by H₂O, instead of DNA-leading toalkylative DNA cleavage. Since the electrophilic center (C-6) of theepisulfonium ion is a 3° carbon in Lnm, the nucleophilic attack of thelatter by H₂O or the —NH₂ group of DNA likely proceeds via a S_(N)1mechanism. Consequently, little discrimination between the twonucleophiles are observed (Asai et al. (1997) Bioorg. Med. Chem. 5:723-729). In contrast, 6-dimethyl-Lnm will generate an episulfonium witha 2° carbon as the electrophilic center. The latter intermediate shouldbe more stable and is likely to alkylate DNA via a S_(N)2 mechanism.Consequently, being a much stronger nucleophile than H₂O, the —NH₂ groupof DNA should be selectively alkylated, leading to DNA cleavage, and H₂Omay be not a stronger enough nucleophile to attack the episulfonium toform the inactive adduct A (at least competed unfavorably in thepresence of DNA). Finally 8-dehydroxyl-6-demethyl-Lnm should exhibitboth an improved stability and selectivity towards DNA cleavage, servingas a good drug candidate (FIG. 8).

[0148] Other preferred embodiments contemplate the synthesis of Lnmanalogs with an oxazole or bithiazole moiety—oxazolyl-Lnm ordithiazolyl-Lnm, respectively (FIG. 7). The former could be prepared byreplacing the cysteine-specific A domain at NRPS-2 with aserine-specific A domain (Shen et al. (1999) Bioorg. Chem., 27:155-171), and the latter could be effected by replacing the NRPS-2module with the bithiazole-forming NRPS modules from the bleomycin genecluster (see PCT/US00/00445). Since it is known that oxazole andthiazole play an important role in drug-DNA recognition (Li et al.(1996) Science 274: 1188-1193; Roy et al. (1999) Nat. Prod. Rep., 16:249-263), it is reasonable to assume that these novel oxazole- orbithiazole-containing Lnm analogs may exhibit improved efficacy asanticancer agents. These five examples are only representatives of thetypes of Lnm analogs that could be prepared by rational engineering ofthe Lnm NRPS/PKS genes, with the choice being influenced byconsiderations of mechanism of action for Lnm. We envisage that variousother permutations can be introduced into Lnm by genetic manipulation ofthe Lnm NRPS and PKS genes.

[0149] Engineering of both NRPS and PKS by either domain or moduleswapping has been very successful for making novel peptides (Stachelhauset al. (1995) Science, 269: 69-72; Belshaw et al. (1999) Science, 284:486-489; Stachelhaus et al. (1999) Chem. Biol. 6: 493-505; de Ferra etal. (1997) J. Biol. Chem., 272: 25304-25309; Elsner et al. (1997) J.Biol. Chem., 272: 4814-4819; Schneider et al. (1998) Mol. Gen. Genet.,257: 308-318; Stachelhaus and Marahiel (1996) Biochem. Pharmacol., 52:177-186) and polyketides (Donadio et al. (1993) Proc. Natl. Acad Sci.,USA, 90: 7119-7123; Cortes et al. (1995) Science, 268: 1487-1489; Kao etal. (1994) Science, 265: 509-512; Kao et al. (1994) J. Am. Chem. Soc.,116: 11612-11613; Kao et al. (1995) J. Am. Chem. Soc., 117: 9105-9106;McDaniel et al. (1997) J. Am. Chem. Soc., 119: 4309-4310; Pieper et al.(1997) Biochemistry, 36: 1846-1851; Bedford et al.(1996) Chem. Biol., 3:826-831; Oliynyk et al. (1996) Chem. Biol., 3: 833-839; McDaniel et al.(1999) Proc. Natl. Acad. Sci., USA, 96: 1846-1851; Jacobsen et al.(1997) Science, 277: 367-369; Gokhale et al. (1999) Science, 284:482-485; Ruan et al. (1997) J. Bacteriol., 179: 6416-6425; Stassi et al.(1998) Proc. Natl. Acad. Sci., USA, 95: 7305-7309) with the desiredstructural alterations. Domain or modules boundaries for both NRPS andPKS are well defined, although the effectiveness of individual domain ormodule swamping experiment is preferably empirically determined.

[0150] In certain preferred embodiments, macrolactam products areisolated and subjected to mass spectrum and 1-D and 2-D NMR analyses todetermine whether inactivation of lnmA, lnmB, lnmZ, and/or the MT domainof PKS-3 in lnmJ, respectively, has resulted in the production of8-deoxyl-LNM, or 6-demethyl-Lnm, respectively. Similar strategy is usedto carry out the double inactivation of the hydroxylase and methyltransferase genes for the production of 8-dehydroxyl-6-demethyl-Lnm, aswell as the desired domain or module replacement of the Lnm NRPS-2 toconstruct S. atroolivaceus recombinant strains producing oxazolyl-Lnmand dithiazoly-Lnm.

[0151] Although in vivo manipulation of Lnm biosynthesis in S.atroolivaceus has been demonstrated as feasible herein, in certainembodiments, methods to clone the entire lnm gene cluster into e.g., S.lividans or S. coelicolor—either by the newly developed E.coli-Streptomyces artificial chromosome (Sosio et al. (2000) NatureBiotechnol. 18: 343-345) or by the multi-plasmid approach (Tang et al.(2000) Science, 87: 640-642; Xue et al. (1999) Proc. Natl. Acad. Sci.USA, 96: 11740-11745) (up to three compatible Streptomyces vectors) areavailable and can be used.

[0152] Production of novel leinamycins by engineering leinamycinbiosynthesis has been demonstrated with lnmH as an example. LnmH is aprotein of unknown function on the basis of amino acid sequenceanalysis. Inactivation of lnmh yields an S. atroolivaceus mutant that nolonger produces leinamycin but accumulates at least two new leinamycinmetabolites upon HPLC analysis (see FIGS. 9). The production of thesenew metabolites results exclusively from the inactivation of ORF43.Complementation of the lnmH mutant by overexpression of lnmH under theermE* promoter in a low-copy-number plasmid restores the leinamycinproduction to the mutant strain with the same metabolite profile as thewild type S. atroolivaceus strain.

[0153] B) Synthesis of Leinamycin Analogs by “Random” Modification oflnm Genes.

[0154] Leinamycin analogs can also be synthesized byrandomly/haphazardly altering genes in the lnm cluster expressing theproducts of the randomly modified megasynthetase and then screening theproducts for the desired activity. Methods of “randomly” altering lnmcluster genes are described below.

[0155] V. Generation of other Synthetic Systems.

[0156] In addition to the production of leinamycin or modifiedleinamycins, the leinamycin gene cluster or elements thereof can be usedby themselves or in combination with NRPS and/or PKS modules and/orenzymatic domains of other PKS and/or NRPS systems to produce a widevariety of compounds including, but not limited to various polyketides,polypeptides, polyketide/polypeptide hybrids, various oxazoles andthiazoles, various sugars, various methylated polypeptides/polyketides,and the like. As with the production of modified leinamycins describedabove, such compounds can be produced, in vivo or in vitro, by catalyticbiosynthesis using large, modular PKSs, NRPSs, and hybrid PKS/NRPSsystems. The megasynthetases directing such syntheses can be rationallydesigned e.g. by predetermined alteration/modification of polyketideand/or polypeptide and/or hybrid PKS/NRPS pathways. Alternatively, largecombinatorial libraries of cells harboring various megasynthetases canbe produced by the random modification of particular pathways and thenselected for the production of a molecule or molecules of interest. Itwill be appreciated that, in certain embodiments, such libraries ofmegasynthetases/modified pathways, can be used to generate large,complex combinatorial libraries of compounds which themselves can bescreened for a desired activity.

[0157] A) Directed Modification of Biomolecules.

[0158] Elements (e.g. open reading frames) of the leinamycinbiosynthetic gene cluster and/or variants thereof can be used in a widevariety of “directed” biosynthetic processes (i.e. where the process isdesigned to modify and/or synthesize one or more particular preselectedmetabolite(s)). Polypeptides encoded by particular open reading framesor combinations of open reading frames can be utilized to performparticular chemical modifications of biological molecules.

[0159] Thus, for example, open reading frames encoding a polypeptidesynthetase can be used to chemically modify an amino acid by coupling itto another amino acid. One of skill in the art, utilizing theinformation provided here, can perform literally countless chemicalmodifications and/or syntheses using either “native” leinamycinbiosynthesis metabolites as the substrate molecule, or other moleculescapable of acting as substrates for the particular enzymes in question.Other substrates can be identified by routine screening. Methods ofscreening enzymes for specific activity against particular substratesare well known to those of skill in the art.

[0160] The biosyntheses can be performed in vivo, e.g. by providing ahost cell comprising the desired leinamycin gene cluster open readingframe(s) and/or in vitro, e.g., by providing the polypeptides encoded bythe leinamycin gene cluster orfs and the appropriate substrates and/orcofactors.

[0161] B) Directed Engineering of Novel Synthetic Pathways.

[0162] In numerous embodiments of this invention, novel polyketides,polypeptides, and combinations thereof are created by modifying knownPKSs or NRPSs so as to introduce variations into known polymerssynthesized by the enzymes. Such variations may be introduced by design,for example to modify a known molecule in a specific way, e.g. byreplacing a single monomeric unit within a polymer with another, therebycreating a derivative molecule of predicted structure. Such variationscan also be made by adding one or more modules to a known PKS or NRPS,or by removing one or more module from a known PKS or NRPS. Such novelPKSs or NRPSs can readily be made using a variety of techniques,including recombinant methods and in vitro synthetic methods.

[0163] Using any of these methods, it is possible to introduce PKSdomains into a NRPS, or vice versa, thereby creating novel moleculesincluding both peptide and polyketide structural domains. For example, aPKS enzyme producing a known polyketide can be modified so as to includean additional module that adds a peptide moiety into the polyketide.Novel molecules synthesized using these methods can be screened, usingstandard methods, for any activity of interest, such as antibioticactivity, effects on the cell cycle, effects on the cytoskeleton, etc.

[0164] Novel polyketides, polypeptides, or combinations thereof can alsobe made by creating novel PKSs or NRPSs de novo, using recombinant or invitro synthetic methods. Such novel arrangements of domains can bedesigned, i.e. to create a specific polymer. In addition to creatingnovel PKSs or NRPSs by combining modules, the methods of this inventioncan also be used to make novel modules that can add new monomeric unitsto a growing polypeptide or polyketide chain. Because the identity ofeach module, and, consequently, the identity of the monomer added by themodule, is determined by the identity and number of the functionaldomains comprising the module, it is possible to produce novel monomericunits by creating novel combinations of functional domains within amodule. Such novel modules can be created by design, for example to makea specific module that will add a specific monomer to a polyketide orpolypeptide, or can be created by the random association of domains soas to produce libraries of novel modules. Such novel modules can be madeusing recombinant or in vitro synthetic means.

[0165] Mutations can be made to the native NRPS and/or PKS subunitsequences and such mutants used in place of the native sequence, so longas the mutants are able to function with other PKS and/or PKS subunitsto collectively catalyze the synthesis of an identifiable polyketideand/or polypeptide. Such mutations can be made to the native sequencesusing conventional techniques such as by preparing syntheticoligonucleotides including the mutations and inserting the mutatedsequence into the gene encoding a NRPS and/or PKS subunit usingrestriction endonuclease digestion. (see, e.g., Kunkel, (1985) Proc.Natl. Acad. Sci. USA 82: 448; Geisselsoder et al. (1987) BioTechniques5: 786). Alternatively, the mutations can be effected using a mismatchedprimer (generally 10-20 nucleotides in length) that hybridizes to thenative nucleotide sequence, at a temperature below the meltingtemperature of the mismatched duplex. The primer can be made specific bykeeping primer length and base composition within relatively narrowlimits and by keeping the mutant base centrally located (Zoller andSmith (1983) Meth, Enzymol. 100: 468). Primer extension is effectedusing DNA polymerase, the product cloned and clones containing themutated DNA, derived by segregation of the primer extended strand,selected. Selection can be accomplished using the mutant primer as ahybridization probe. The technique is also applicable for generatingmultiple point mutations (see, e.g., Dalbie-McFarland et al. (1982)Proc. Natl. Acad. Sci USA 79: 6409). PCR mutagenesis will also find usefor effecting the desired mutations.

[0166] C) Random Modification of PKS/NRPS Pathways.

[0167] In another embodiment, variations can be made randomly, forexample by making a library of molecular variants of a known polymer byrandomly mutating one or more PKS or NRPS modules and/or enzymaticdomains or by randomly replacing one or more modules or enzymaticdomains in a known PKS or NRPS with a collection of alternative modulesand/or enzymatic domains.

[0168] The PKS and/or NRPS modules can be combined into a singlemulti-modular enzyme, thereby dramatically increasing the number ofpossible combinations obtained using these methods. These combinationscan be made using standard recombinant or nucleic acid amplificationmethods, for example by shuffling nucleic acid sequences encodingvarious modules or enzymatic domains to create novel arrangements of thesequences, analogous to DNA shuffling methods described in Crameri etal., (1998) Nature 391: 288-291, and in U.S. Pat. Nos. 5,605,793 and in5,837,458. In addition, novel combinations can be made in vitro, forexample by combinatorial synthetic methods. Novel polymers, or polymerlibraries, can be screened for any specific activity using standardmethods.

[0169] Random mutagenesis of the nucleotide sequences obtained asdescribed above can be accomplished by several different techniquesknown in the art, such as by altering sequences within restrictionendonuclease sites, inserting an oligonucleotide linker randomly into aplasmid, by irradiation with X-rays or ultraviolet light, byincorporating incorrect nucleotides during in vitro DNA synthesis, byerror-prone PCR mutagenesis, by preparing synthetic mutants or bydamaging plasmid DNA in vitro with chemicals. Chemical mutagens include,for example, sodium bisulfite, nitrous acid, hydroxylamine, agents whichdamage or remove bases thereby preventing normal base-pairing such ashydrazine or formic acid, analogues of nucleotide precursors such asnitrosoguanidine, 5-bromouracil, 2-aminopurine, or acridineintercalating agents such as proflavine, acriflavine, quinacrine, andthe like. Generally, plasmid DNA or DNA fragments are treated withchemicals, transformed into E. coli and propagated as a pool or libraryof mutant plasmids.

[0170] Large populations of random enzyme variants can be constructed invivo using “recombination-enhanced mutagenesis.” This method employs twoor more pools of, for example, 10⁶ mutants each of the wild-typeencoding nucleotide sequence that are generated using any convenientmutagenesis technique, described more fully above, and then insertedinto cloning vectors.

[0171] D) Incorporation and/or Modification of Non-lnm Cluster Elements.

[0172] In either the directed or random approaches, nucleic acidencoding novel combinations of modules and/or enzymatic are introducedinto a cell. In one embodiment, nucleic acids encoding one or more PKSor NRPS domains are introduced into a cell so as to replace one or moredomains of an endogenous PKS or NRPS within a chromosome of the cell.Endogenous gene replacement can be accomplished using standard methods,such as homologous recombination. Nucleic acids encoding an entire PKS,NRPS, or combination thereof can also be introduced into a cell so as toenable the cell to produce the novel enzyme, and, consequently,synthesize the novel polymer. In a preferred embodiment, such nucleicacids are introduced into the cell optionally along with a number ofadditional genes, together called a ‘gene cluster,’ that influence theexpression of the genes, survival of the expressing cells, etc. In aparticularly preferred embodiment, such cells do not have any other PKS-or NRPS-encoding genes or gene clusters, thereby allowing thestraightforward isolation of the polymer synthesized by the genesintroduced into the cell.

[0173] Furthermore, the recombinant vector(s) can include genes from asingle PKS and/or NRPS gene cluster, or may comprise hybrid replacementPKS gene clusters with, e.g., a gene for one cluster replaced by thecorresponding gene from another gene cluster. For example, it has beenfound that ACPs are readily interchangeable among different synthaseswithout an effect on product structure. Furthermore, a given KR canrecognize and reduce polyketide chains of different chain lengths.Accordingly, these genes are freely interchangeable in the constructsdescribed herein. Thus, the replacement clusters of the presentinvention can be derived from any combination of PKS and/or NRPS genesets that ultimately function to produce an identifiable polyketideand/or peptide.

[0174] Examples of hybrid replacement clusters include, but are notlimited to, clusters with genes derived from two or more of the act genecluster, the whiE gene cluster, frenolicin (fren), granaticin (gra),tetracenomycin (tcm), 6-methylsalicylic acid (6-msas), oxytetracycline(otc), tetracycline (tet), erythromycin (ery), griseusin (gris),nanamycin, medermycin, daunroubicin, tylosin, carbomycin, spiramycin,avermectin, monensin, nonactin, curamycin, rifamycin and candicidinsynthase gene clusters, among others. (For a discussion of various PKSs,see, e.g., Hopwood and Sherman (199) Ann. Rev. Genet. 24: 37-66; O'Hagan(1991) The Polyketide Metabolites, Ellis Horwood Limited.

[0175] A number of hybrid gene clusters have been constructed, havingcomponents derived from the act, fren, tcm, gris and gra gene clusters(see, e.g., U.S. Pat. No. 5,712,146). Other hybrid gene clusters, asdescribed above, can easily be produced and screened using thedisclosure herein, for the production of identifiable polyketides,polypeptides or polyketide/polypeptide hybrids.

[0176] Host cells (e.g. Streptomyces) can be transformed with one ormore vectors, collectively encoding a functional PKS/NRPS set (e.g. aleinamycin or leinamycin analog), or a cocktail comprising a randomassortment of PKS and/or NRPS genes, modules, active sites, or portionsthereof. The vector(s) can include native or hybrid combinations of PKSand/or NRPS subunits or cocktail components, or mutants thereof. Asexplained above, the gene cluster need not correspond to the completenative gene cluster but need only encode the necessary PKS and/or NRPScomponents to catalyze the production of the desired product. Forexample, in Streptomyces aromatic PKSs, carbon chain assembly requiresthe products of three open reading frames (ORFs). ORF1 encodes aketosynthase (KS) and an acyltransferase (AT) active site (KS/AT); ORF2encodes a chain length-determining factor (CLF), a protein similar tothe ORF1 product but lacking the KS and AT motifs; and ORF3 encodes adiscrete acyl carrier protein (ACP). Some gene clusters also code for aketoreductase (KR) and a cyclase, involved in cyclization of the nascentpolyketide backbone. However, it has been found that only the KS/AT,CLF, and ACP, need be present in order to produce an identifiablepolyketide. Thus, in the case of aromatic PKSs derived fromStreptomyces, these three genes, without the other components of thenative clusters, can be included in one or more recombinant vectors, toconstitute a “minimal” replacement PKS gene cluster.

[0177] E) Variation of Starter and Extender Units.

[0178] In addition to varying the PKS and/or NRPS modules and/ordomains, variations in the products produced by various PKS/NRPS systemscan be obtained by varying the starter units and/or the extender units.Thus, for example, a considerable degree of variability exists forstarter units, e.g., acetyl CoA, malonyl CoA, propionyl CoA, acetate,butyrate, isobutyrate and the like. In addition, naturally occurringPKSs and/or NRPSs have shown some tolerance for varying extender units.

[0179] F) Screening of Products.

[0180] Particularly where large combinatorial libraries are synthesized,e.g. using one or more modules and/or enzymatic domains of the lnm genecluster it will often be desired to screen the resulting compound(s) forthe desired activity. Methods of screening compounds (e.g. polypeptides,polyketides, sugars, thiazoles, etc.) for various activities of interest(e.g cytotoxicity, antimicrobial activity, particular chemicalactivities, etc.) are well known to those of skill in the art.

[0181] Where large numbers of compounds are produced, it is oftendesired to rapidly screen such compounds using “high throughput systems”(HTS). High throughput assays systems are well known to those of skillin the art and many such systems are commercially available. (see, e.g.,Zymark Corp., Hopkinton, Mass.; Air Technical Industries, Mentor, Ohio;Beckman Instruments, Inc. Fullerton, Calif.; Precision Systems, Inc.,Natick, Mass., etc.). These systems typically automate entire proceduresincluding all sample and reagent pipetting, liquid dispensing, timedincubations, and final readings of the microplate(s) in detector(s)appropriate for the assay. These configurable systems provide highthroughput and rapid start up as well as a high degree of flexibilityand customization. The manufacturers of such systems typically providedetailed protocols for the various high throughput screens.

[0182] VI. In vitro Syntheses.

[0183] In additional embodiments of this invention, leinamycins andother polyketides and/or polypeptides are synthesized and/or modified invitro. Individual enzymatic domains or modules can be used in vitro tomodify a unit and/or to add a single monomeric unit to a growingpolyketide or polypeptide chain. In one approach a metasynthetaseproviding all the desired synthetic activities recombinantly expressedand then provided, the appropriate substrates and buffer system e.g. ina bioreactor, to direct the synthesis of the desired product. In anotherapproach, various PKSs and/or NRPSs are provided in different solutionsand the growing polymer chains can be sequentially introduced into theplurality of solutions, each containing a single (or several) PKS orNRPS modules. In still another embodiment, the PKS and/or NRPS modulesor enzymatic domains are provided attached to a solid support and afluid containing the growing macromolecule is passed over the surfacewhereby the PKSs or NRPSs are able to react with the target substrate.

[0184] In one preferred embodiment, a combinatorial library ofpolyketides or polypeptides, or combinations thereof, is created byusing automated means to facilitate the sequential introduction of amultitude of polymeric chains, each attached to a solid support, to acollection of solutions, each containing a single PKS or NRPS module.These automated means can be used to systematically vary the sequence bywhich each polymeric chain is introduced into the various solutions,thereby creating a combinatorial library. Numerous methods are wellknown in the art to create combinatorial libraries of molecules by thesequential addition of monomeric units, for example as described in WO97/02358.

[0185] VII. Kits.

[0186] In still another embodiment, this invention provides kits forpractice of the methods described herein. In one preferred embodiment,the kits comprise one or more containers containing nucleic acidsencoding one or more of the lnm gene cluster ORFs and/or one or more ofthe lnm PKS or NRPS modules or enzymatic domains. Certain kits maycomprise vectors encoding the lnm orfs and/or cells containing suchvectors. The kits may optionally include any reagents and/or apparatusto facilitate practice of the assays described herein. Such reagentsinclude, but are not limited to buffers, labels, labeled antibodies,bioreactors, cells, etc.

[0187] In addition, the kits may include instructional materialscontaining directions (i.e., protocols) for the practice of the methodsof this invention. Preferred instructional materials provide protocolsutilizing the kit contents for creating or modifying lnm module or ORFand/or for synthesizing or modifying a molecule using one or more lnmmodules and/or enzymatic domains. While the instructional materialstypically comprise written or printed materials they are not limited tosuch. Any medium capable of storing such instructions and communicatingthem to an end user is contemplated by this invention. Such mediainclude, but are not limited to electronic storage media (e.g., magneticdiscs, tapes, cartridges, chips), optical media (e.g., CD ROM), and thelike. Such media may include addresses to internet sites that providesuch instructional materials.

[0188] VIII. Methods of Utilizing LnmG and Discrete Acyltransferasesfrom Related Type I PKS and Type I PKS/NRPS Systems.

[0189] The majority of type I PKSs characterized to date include modulesminimally containing three domains, β-ketoacyl synthase (KS), acyltransferase (AT), and acyl carrier protein (ACP), that select, activate,and catalyze a Claisen condensation between the extender unit and thegrowing polyketide chain from the proceeding module, generating aβ-ketoacyl-S-ACP intermediate. Optional domains are found between AT andACP that carry out the variable set of reductive modifications of theβ-keto group before the next round of chain extension.

[0190] In modular PKS systems, selection of the extender unit for eachmodule, as well as the starter unit in many cases, is carried out by theAT domain (Reeves et al, Biochemistry 2001, 40, 15464-15470). Extenderand starter molecules include (e.g., malonyl CoA, alkyl malonyl CoA(including methyl malonyl CoA, ethyl malonyl CoA and propionyl malonylCoA), acyl malonyl CoA, hydroxy malonyl CoA and alkoxy malonyl CoA(e.g., methoxy malonyl CoA). An AT domain catalyzes the transacylationof the monomer unit from CoA to the phosphopantetheine arm of the acylcarrier protein (ACP) in the same module. AT domains generally possess astringent specificity for a single acyl-CoA substrate in their naturalcontext, although some ATs can incorporate at least two differentmonomers with similar efficiencies. Amino acid sequence alignmentsbetween methylmalonyl-CoA (mmCoA)-specific and malonyl-CoA(mCoA)-specific AT domains cluster into two groups according to thespecificity of the domain (Schwecke et al (1995) Proc. Natl. Acad. Sci.USA 92, 7839-7843), Kakavas et al (1997) J. Bacteriol. 179, 7515-7522.and Wu et al (2000) Gene 251, 81-90). At least three divergent sequencemotifs for AT domains have been identified on the basis of suchalignments and comparison to the Escherichia coli malonyl-CoA:ACPtransacylase (FabD) crystal structure (Haydock, et al. (1995) FEBS Lett.374, 246-248, and Serre et al. (1995) J. Biol. Chem. 270, 12961-12964and Ikeda et al., Proc. Natl. Acad. Sci. USA, 96, 9509-9514).

[0191] Despite the identification of regions within modular ATs thatcorrelate with specificity, the most popular method of engineeringsubstrate utilization in PKSs has been via the exchange of the entire˜300-350-amino acid AT domain of one module with a homologous AT domainspecific for a different starter or extender unit, obtained from anothermodule, usually of a heterologous PKS. A number of successful ATreplacements with modular PKSs have been reported, in most caseproducing a new polyketide with the predicted structural change (Kuhtosset al. (1996) Gene 183, 231 -236, Oliynyk et al. (1996) Chem. Biol. 3,833-839, Liu et al. (1997) J. Am. Chem. Soc. 119, 10553-10554, and Ruanet al. (1997) J. Bacteriol. 179, 6416-6425, Stassi et al. (1998) Proc.Natl. Acad. Sci. USA 95, 7305-7309 and McDaniel et al. (1999) Proc.Natl. Acad. Sci. USA 96, 1846-1851)(e.g., malonyl CoA, methyl malonylCoA, ethyl malonyl CoA, acyl malonyl CoA, hydroxy malonyl CoA andmethoxy malonyl CoA). McDaniel et al. constructed a combinatoriallibrary of polyketides using 6-ethoxyerythronolide B synthase (DEBS),the PKS that produces the macrolide ring of erythromycin. This wasaccomplished by substituting the modular ATs and beta-carbon processingdomains of DEBS with counterparts from rapamycin PKS (RAPS) that encodealternative substrate specificities and beta-carbonreduction/dehydration activities. However, occasionally AT replacementsare only marginally successful or entirely unsuccessful, leading to onlyvery small amounts of the desired compound or no product at all, forunknown reasons (Ruan et al. (1997) J. Bacteriol. 179, 6416-6425).

[0192] If it is assumed that the genetic engineering of the PKS resultsin little or no predicted product, there are at least two possiblereasons for such failures. Either the foreign AT domain causes the PKScomplex to fold incorrectly and lose a necessary activity, or thereplacement leads to a modified polyketide chain that is not recognizedas a substrate by a subsequent (“downstream”) PKS activity. In theformer case, success might be achieved by changing AT domain specificityin a manner that minimized perturbation to the tertiary structure of themodule, e.g., mutagenesis of a limited number of amino acids. To do so,however, requires an understanding of the complex rules for proper PKSfolding and the structural features involved in substrate recognition.

[0193] The inventors' characterization of the leinamycin biosynthesisgene cluster has revealed that the leinamycin (LNM) PKS fromStreptomyces alroolivaceus S-140 is composed of six modules, all ofwhich contain the core KS and ACP domains as well as the variable set ofoptional domains but completely lack a cognate AT domain indicative of atype I PKS. Instead, the inventors identified that a discrete,interactive AT protein, acting in trans, loads the malonyl CoA extenderunit onto the ACPs of all six PKS modules. This finding provides newinsights into PKS structure and mechanism and suggests an alternativemodel for a type I PKS in which the KS and ACP domains of each modulecould minimally constitute the core structure. This structure furthersuggests a novel way in which PKS and PKS/NRPS systems may beadvantageously manipulated without the disadvantages of the engineeringapproaches described above.

[0194] Rare exceptions to the general type I PKS model have also beenobserved in bacterial PKS systems which may possess separate AT enzymesencoded elsewhere within their genomes to act in trans to load theappropriate acyl groups (Duitman et al., 1999; Paitan et al., 1999;Huang et al., 2001. Zhu et al. 2002, Gene 298: 79-89). A PKS containingsuch putative “super ATs” is the only partly described pks cluster(Albertini et al. (1995) Microbiology 141, 299-309) in the genome ofBacillus subtilis. Its gene products consist of a large number of PKSmodules without AT domains and, encoded at the upstream end of thecluster, three isolated ATs. The secondary metabolite generated by theseproteins is not known. Another related system is the mupirocin PKS fromPseudomonas fluorescens recently submitted to Gen-Bank (accession no.AF318063). This cluster contains a single gene with two AT domains. Asmall number of further PKS modules lacking AT domains are known fromthe albicidin (Huang et al, (2001) Microbiology 147, 631-642) and TAbiosynthesis gene clusters (Paitan et al. (1999) J. Mol. Bio. 286,465-474). Both systems have only been analyzed in part thus far, but itis very likely that they also contain discrete AT genes.

[0195] Also, AT homologs are completely absent in the PKS/NRPSresponsible for pederin production in the bacterial symbiont of Paederusbeetles. (Peil, J. (2002) Proc. Natl. Acad. Sci., 99: 14002-14007.) Thepederin system's alignment with other known type I PKSs revealed that acontinuous ≈300-aa region of each AT domain, including the active-siteGHS motif, are deleted in the type I PKSs PedF and PedG, with no otherdomain replacing them. PedC and PedD, reside upstream of pedF and appearto be discrete AT enzymes.

[0196] Data supportive of the trans loading of the Lnm PKS by LnmG, adiscrete transacylase (AT), is set forth in Example 2. Embodiments ofthe present invention therefore include the use of the discrete AT, LnmGor a catalytically active fragment thereof, in a wide variety ofbiosynthetic processes wherein the acyltransferase activity of LnmG isutilized to load extender molecules onto ACP domains of PKS modules.Furthermore, the present invention is also directed to LnmG-likemolecules, homologs of LnmG, or fragments thereof which can be utilizedin similar fashion to LnmG in the following methods.

[0197] Processes utilizing discrete LnmG include “directed” approaches(i.e., where the process is designed to modify and/or synthesize one ormore particular preselected metabolites(s)). For example, using knownrecombinant or in vitro methods, LnmG may be substituted for a differentnaturally occurring discrete AT in a non-leinamycin biosynthesis systemthat relies, at least partially, on a discrete AT for loading ofextender and starter molecules on PKS modules.

[0198] This approach facilitates the selective introduction ofparticular extender molecules at selected positions within a productbased on a limited knowledge of the modular structure of the PKS and theextender molecule specificity of the substituted discrete AT. Forexample, using recombinant methods known in the art, a naturallyoccurring discrete AT associated with a type I polyketide system andspecific for methyl malonyl CoA may be substituted with an alternativediscrete AT, preferably LnmG. Consequently, malonyl CoA extendersprovided by the substituted LnmG may be utilized by a type I PKS modulethat naturally utilizes methyl malonyl CoA resulting in the productionof a novel product with a predicted structure.

[0199] In a further embodiment of the present invention, a modular ATdomain contained within a PKS module may be unnaturally renderedineffective (i.e., deleted or inactivated by recombinant or in vitromethods) to provide a PKS module lacking the ability to itself load anextender molecule onto its ACP domain. Subsequently, a discrete ATacting in trans, such as LmnG, could “rescue” the inactivated module andafford the effective loading of extender molecule on the PKS module sothat a chain elongation step may occur. This approach is particularlyadvantageous over prior domain “swapping” approaches where an AT domainis deleted from a PKS module and a second AT domain is substituted intothe modular structure creating the opportunity for protein mis-folding,etc. Thus, for example, a site-directed mutation to key residuesnecessary for acyltransferase catalytic activity of a modular AT domainmay be made by standard techniques and the AT domain, in inactivatedform, allowed to reside in the modular PKS. The acyltransferase activitynecessary to load an extender molecule onto the ACP of the “inactivated”PKS module could then be supplied in trans by a discrete AT, preferablya discrete AT having specificity for a different extender molecule thanthe inactivated AT domain (e.g., a discrete AT with specificity formalonyl CoA, such as LnmG, could substitute for an inactivated modularAT domain with specificity for a methyl malonyl CoA extender; thus, amalonate unit would be incorporated into the product structure insteadof a methyl malonate unit).

[0200] In a host cell, the discrete AT could be, for example, providedon a vector comprising a nucleic acid encoding at least a catalyticdomain of a discrete AT. In addition, the PKS module may be provided ona different vector including a nucleic acid encoding the PKS module.Thus, a two plasmid heterologous system could be practiced wherein thedifferent vector comprising at least the PKS module may further includecoding sequence for additional PKS modules, preferably, modulessufficient to produce a predicted product.

[0201] In an in vitro embodiment, the discrete AT may be provided as arecombinantly expressed or purified version of a discrete AT provided ina addition to various isolated domains of a PKS or PKS/NRPS, includingat least the “inactivated” PKS module requiring trans activity to loadextender molecules. Thus, novel polyketides and polyketide/peptideproducts may be produced by the novel combination of discrete AT andmodified PKS modules harboring “inactivated” modular AT domains whereinthe discrete AT domain has a different extender molecule specificitythan the “inactivated” AT domain.

[0202] A specific example meant for illustration purposes only, may behad by reference to the production of the natural polyketide6-deoxyerythronolide B (6-dEB) by the type I PKS 6-deoxyerythronolide Bsynthase (DEBS). McDaniel et al. previously described the substitutionof an AT cassette from module 2 of the rapamycin PKS to alter theextender unit specificity from methyl malonate CoA to malonyl CoA inmodules 1-3, 5, and 6 of DEBS. (McDaniel et al., (1999) Proc. Natl.Acad. Sci. U.S.A. 96: 1846-1851.) However, substitution at module 4 ofDEBS proved unsuccessful. Subsequently, Reeves et al. identified apossible route around the module 4 problem by site-directed mutagenesisof specific residues within module 4 to alter the module's extenderspecificity from methyl malonate to malonate. (See, e.g., Reeves et al.(2001) Biochemistry 40:15464-15470.) However, domain swapping (McDanielet al. (1999)) and the alternative of site-directed mutagenesis (Reeveset al. (2001)) key substrate specificity residues are labor intensiveapproaches requiring extensive knowledge of sequence and structure ofthe modular AT domain.

[0203] In this regard, the present invention may be utilized tocircumvent the disadvantages of the previous methods. For example, thespecificity of DEBS modules 1-6, taken individually, in combination, orin their entirety may be had by inactivation of the well known catalyticresidues (e.g., by site-directed mutagenesis) responsible for acyltransferase activity in ATs followed by providing a discrete AT, such asLnmG, to supply, in trans, the required acyl transferase activity forthe “inactivated” module. As described above, a two plasmid approach ina transformed host cell could be utilized wherein LnmG is encoded by afirst plasmid and the respective modified module(s) of DEBs encoded by asecond, different plasmid. In the case of DEBS, LnmG would provide asubstitution of a malonate unit for a methyl malonate unit and result inthe production of 6-dEB analogs. For example, inactivation of themodular AT domain of DEBS module 4 and substitution of the LnmG acyltransferase activity, in trans, would result in the production of6-desmethyl-6-deoxyerythronolide B (6-desmethyl-6-dEB), a valuableanalog of the natural polyketide 6-dEB. Thus, the large-scale remodelingof a modular PKS is not necessary nor is extensive knowledge regardingthe complexities of substrate specificity in the present invention.

[0204] In another embodiment of the invention, an in vitro method isprovided for loading an ACP isolated from a type I PKS with an extendermolecule by utilizing the trans acyltransferase activity of a discreteAT, preferably LnmG. Such an approach has been reduced to practice bythe present inventors, as described in Example 2, such that holo-ACPdomains may be loaded with extender molecule in vitro by the discrete ATLmnG. Such an approach facilitates ease of assembly of directedbiosythethic systems, particularly in vitro systems where PKS orPKS/NRPS domains/modules may be specifically selected and combined inreactions in whatever order selected by the worker. For example, aholo-ACP domain may be incubated with an extender molecule and aspecific discrete AT selected for its extender molecule specificityunder conditions sufficient to load the extender molecule onto theholo-ACP domain. In a subsequent reaction, the loaded holo-ACP domainmay be contacted with a PKS domain to facilitate the starting step ofpolyketide chain elongation or a further chain elongation step whereinthe PKS domain carries a nascent polyketide elongation product to becoupled to the extender molecule. General discussions of in vitrosynthesis are provided in Section VII above.

[0205] Discrete ATs may also be substituted for modular AT domains intype I PKS or type I PKS/NRPS according to domain swapping strategieswell know in the field, and previously described herein (e.g.,recombinant methods and in vitro synthetic methods). Domain swapping maybe particularly attractive in directed biosynthesis of a predictedpolyketide product wherein the desired product necessitates the use ofPKS domains with elongation activities utilizing particular extender orstarter molecules. As a general example, a type I PKS module having anAT domain specific for loading of malonyl CoA may be altered by thesubstitution of an AT domain having extender molecule specificity formethoxy malonyl CoA. It is envisioned that, for example, a discrete ATmay be, instead of utilized in a trans manner, substituted into themodular structure of a type I PKS or type I PKS/NRPS to provideacyltransferase activity in a noniterative manner, in similar fashion tothe wide variety of naturally-occurring type PKS having noniterative ATdomains within their modular structure. Such domain swapping has beenpreviously effective to produce novel polyketides and/orpolyketide/peptide hybrid products. (See, e.g., McDaniel et al. (1999))Combining a discrete AT into a PKS or PKS/NRPS has advantages wheresimplicity in a biosynthetic system is desired, in particular, where asingle gene cluster unit simplifies transformation, maintenance oftransformed cells, and stable expression of biosynthetic products.

[0206] The discovery by the present inventors, that LnmI and LnmJ aretype I PKSs consisting minimally of the KS and ACP domains but lackingthe cognate AT domain and that LnmG is a discrete AT enzyme thatinteracts with LnmIJ to form functional type I PKS modules by providingthe AT activity in trans, unveils a new architecture and mechanism fortype I PKSs and suggest an alternative model for type I PKS with KS andACP as the minimal core structure. Preliminary phylogenetic analysisindeed has led to the identification of LnmG homologs (FIGS. 14 and 15),which, in a mechanistic analogy to LnmG, could provide the AT activityin trans to the PKS modules for polyketide biosynthesis. Thus, LnmGhomologs and/or LnmG-like ATs, no matter which type I PKS or type IPKS/NRPS system initially isolated from, could be utilized in similarfashion to the LnmG-based embodiments set forth above.

[0207] Phylogenetic analysis of LnmG AT and its homologs from other“AT-less” type I PKS clusters and their relationships to cognate ATsfrom type I PKS clusters are shown in FIGS. 14 and 15. The LnmG AT andits homologs fall into distinct groups that differ from cognate ATs oftype I PKSs. Multiple sequence alignment and phylogenetic analysis wereperformed by the CLUSTAL W program (version 1.81), using Gonnet seriesweight matrix with a gap open penalty of 10.00 and a gap extensionpenalty of 0.2 [Higgins, D. G., Thompson, J. D. and Gibson, T. J. (1996)Using CLUSTAL for multiple sequence alignments. Methods Enzymol., 266,383-402]. We predict that PksC/PksD/PksE-AT (for the unknown polyketidein Bacillus subtilis (Albertini, A. M., Caramori, T., Scoffone, F.,Scotti, C., & Galizzi, A. (1995) Microbiology 141, 299-309)), PedC/PedD(for pederin in a bacterial symbiont of Paederus beetles (Piel, J.(2002) Proc. Natl. Acad. Sci. USA 98, 14808-14813)), MmpIII-AT1/AT2 (formupirocin in Pseudomonas fluorescens (accession number AF318063)), FenF(for mycosubtilin in Bacillus subtilis ATCC6633 (Duitman, E. H. et al.(1999) Proc. Natl. Acad. Sci. USA 96, 13294-13299)), and Mx-TaK (for Ta1in Myxococcus xanthus (Paitan, Y., Alon, G., Orr, E., Ron, E. Z., &Rosenberg, E. (1999) J. Mol. Biol. 286, 465-474.)), acting in amechanistic analogy to LnmG (Cheng, Y.-Q., Tang, G.-L., & Shen, B.(2002) J. Bacteriol. 184, 7013-7024), load the malonyl CoA extender unitonto the AT-less PKS modules in trans for polyketide biosynthesis inthese clusters. For cognate ATs from rifamycin (Rif), rapamycin (Rap),erythromycin (Ery), and epothilone (Epo) clusters, protein accessionnumbers are given after the protein names: Rif-AT4 and Rif-AT2, AF04570;Rap-AT1 and Rap-AT2, X86780; Ery-AT1, Q03131; Epo-AT3, AF217189.Abbreviations are: MCoA for malonyl CoA and mMCoA for methyl malonylCoA.

[0208] The distribution in the phylogenic tree suggests that thediscrete AT proteins associated with certain type I PKSs areevolutionarily distant from the cognate AT domains of other type I PKSs.Therefor, LmnG represents a member of a discrete AT family quitedistinct from the cognate AT domains of known type I PKSs. Proteinswithin the bracket in FIG. 14 are anticipated to be LnmG homologs.LnmG-like characteristics include the following properties: (1) the ATis encoded by a discrete gene physically set apart from the sequencesencoding the modules of the respective type I PKS or type I NRPS-PKSwithout at least one cognate AT domain; and (2) the AT provides an acyltransferase activity in trans to the associated type I PKS or type INRPS-PKS lacking at least one cognate AT domain. For the purposes ofpracticing the present invention, LnmG homologs are considered theequivalent of LnmG and may be substituted for LnmG in the variousmethods described above and claimed below, unless noted otherwise.

EXAMPLES

[0209] The following examples are offered to illustrate, but not tolimit the claimed invention.

Example 1 The Biosynthetic Gene Cluster for the Antitumor MacrolactamLeinamycin from Streptomyces atroolivaceus: a Novel Approach forIdentifying and Cloning Thiazole Biosynthetic Genes

[0210] In this example, we describe (1) the construction of a cosmidlibrary of S. atroolivaceus, (2) PCR amplification of a conservedcyclase-domain probe, (3) identification and mapping of overlappingcosmid clones that cover the target Lnm biosynthetic gene cluster andflanking regions, (4) the purification, HPLC and mass spectral (MS)analyses of Lnm production in S. atrooliveceus, (5) the sequencing andsequence analysis of a 11 kb DNA from the gene cluster, (6) thedevelopment of a genetic system, and (7) the confirmation of cloned genecluster encoding Lnm biosyntheses by gene disruption to generate Lnmnon-producing mutants.

[0211] Materials and Methods

[0212] Genomic DNA Isolation.

[0213] Standard protocols are followed in this study (Kieser et al.(2000) Practical Streptomyces genetics. The John Innes Foundation,Norwich, England; Sambrook et al. (1989) Molecular Cloning, A LaboratoryManual (2nd ed.). Cold Spring Harbor Laboratory Press).

[0214] Cosmid Library Construction.

[0215] (1) Cosmid vector pOJ446: The ready-for-use pOJ446 was from labstock (Liu and Shen (2000) Antimicrob Agents Chemother. 44: 382-392;Smith et al. (2000) Antimicrob Agents Chemother. 44: 1809-1817; Du etal. (2000) Chem. Biol. 7: 623-642). The sample contains theHpa-digested, dephosphorylated, and then BamHI-digested pOJ446, i.e. themixture of a ˜2 kb arm and a ˜8 kb arm.

[0216] (2) Preparation of partially digested DNA inserts: Genomic DNA ofS. atroolivaceus was partially digested by MboI (New England Biolabs,MA) following the standard protocol (Rao et al. (1987) Methods inEnzymol. 153: 166-198; Sambrook et al., supra). A total of 50 μg of DNAwas diluted to 900 μl with 10 mM Tris-HCl (pH 8.0) buffer, and 100 μl of10× buffer was added. Let the DNA solution sit on ice for at least 2hrs. Label 10 1.5-ml eppendorf tubes and keep on ice. Aliquot 40 μl ofDNA solution to tube #1; aliquot 20 μl of DNA solution to the rest tubes(#2 to #10). Ten units (1 μl) of MboI was added to tube #1 and mixedwell on ice. Twenty μl of mixture #1 was transferred to tube #2 andmixed. Twenty μl of mixture #2 was transferred to tube #3 and mixed, andso on until tube #9. Discard 20 μl of mixture #9. No enzyme was added totube #10. Incubate the 10 reaction mixtures in 37° C. water bath for 30min. Stop reaction by adding 2 μl of 0.2 M EDTA (pH 8.0; finalconcentration 5 mM). Heat reactions at 70° C. for 15 min to inactivateenzyme. Add 4 μl 6× DNA loading buffer to each tube and run 10 μl ofeach sample in a 0.3% gel. Use intact λ DNA as control. Run gel slowlyfor at least 4 hrs. Use the optimal conditions for large-scale DNApartial digestion as follow: 30 μg DNA with 0.6 unit of MboI in 300 μlvolume. Partially digested DNA solution was diluted with equal volume ofddH2O and extracted once with phenol:chloroform, once with chloroform,and precipitated ethanol. Redissolve DNA in 180 μl ddH2O and treatedwith 10 units of CIP (NEB) at 37° C. for 4 hrs. Perform again phenol:Chloroform extraction and ethanol precipitation. Finally redissolve DNAin 10 μl ddH₂O (˜2 μl/ul).

[0217] (3) Vector-insert ligation: Five μl of MboI partially digested,dephosphorylated S. atroolivaceus DNA fragment (˜10 μg) was ligated with2 μl of HpaI-cut, dephosphorylated and BamHI digested pOJ446 (˜4 μg) inthe volume of 10 μl with 100 units of T4 DNA ligase (NEB) at 16° C. forovernight.

[0218] (4) Packaging for the ligation mixture: Four μl of ligationmixture was packaged with one vial of Gigapack III XL extract(Stratagene, CA) according to manufacturer's instruction. Five hundredμl of SM buffer containing the packaged phage particles was obtained andstored at 4° C.

[0219] (5) Determination of the titer of the cosmid library and libraryamplification: Fresh host cell E. coli XL1-Blue MRF′ suspension in 10 mMMgSO4 was prepared according to manufacture's instruction (Stratagene).Twenty five μl of 1:10 and 1:50 diluted phage particle solutions weremixed with 25 μl of host cells (OD₆₀₀=0.5) and incubated at 37° C. for20 min. Four hundred fifty μl of LB medium was added to each sample andincubated at 37° C. for 1 hr with gently shaking. Aliquots (50 μl, 450μl) of each sample were spread onto [o] 150 mm LB plates (with 100 μg/mlapramycin). Plates were incubated at 37° C. overnight. The number ofcolonies was counted and the titer of the cosmid library was calculated.Cosmid DNA was prepared from 16 random clones, digested with BamHI andfragments were separated in a 0.8% gel to test the quality of library.An aliquot of the primary phage stock equivalent to 3 times of thegenome size of S. atroolivaceus was transducted into XL1-Blue MRF′ cellsand grown on LB plates with 100 μl/ml apramycin. Cells of the colonieswere collected into LB medium with apramycin. Sterile glycerol was addedto a final concentration of 20% and stored at −80° C. as the permanentcosmid library stock.

[0220] Degenerate PCR Primers.

[0221] Primer set for cyclase-domain amplification were: CyFP(5′-GCGCCACGAGCCGTTYCCNYTNAC-3′, SEQ ID NO:215) and CyRP(5′-GCCCAGGTTGGAGGTGARSACNACNGG-3′, SEQ ID NO:216); primer set foroxidase-domain amplification were: OxFP(5′-GCCGGCTCCACCTACCCNGTNCARAC-3′, SEQ ID NO:217) and OXRP(5′-CATCAGCAGCTGGGTCATGKMNCCNGCYTC-3′, SEQ ID NO:218). These primerswere designed by the “Consensus-Degenerate Hybrid OligonucleotidePrimer” strategy (Rose et al. (1998) Nucleic Acids Res. 26: 1628-1635)and synthesized by campus oligo syntheses facility (UC-Davs, CA). Primerset for PKS were: PKSFP (5′-GCSTCCCGSGACCTGGGCTTCGACTC-3′, SEQ IDNO:219) and PKSRP (5′-AGSGASGASGAGCAGGCGGTSTCSAC-3′, SEQ ID NO:220);primer set for PTS were: PTSFP(5′-ATCTACACSTCSGGCACSACSGGCAAGCCSAAGGG-3′, SEQ ID NO:2211) and PTSRP(5′-AWTGAGKSICCICCSRRGIMGAAGAA-3′, SEQ ID NO:222). These primers wereobtained from lab stock.

[0222] PCR Amplification

[0223] All PCR reactions were performed on a GeneAmp 2400 (Perkin Elmer,CA) thermocycler with touchdown programs. A typical PCR reaction (50 μlvolume) contains of 1× PCR buffer with 1.5 mM MgCl₂, 5 ng of template,7.5% DMSO, 100 nM dNTPs, 25 pmol each primer, 2.5 units of Tag DNApolymerase (Boehringer Mannheim Biochemicals, IN). For cyclase-domainamplification, the program was: pre-run denaturation (4 min at 94°C.)→10 cycles of ramp amplification (45 sec denaturation at 94° C.→1 minannealing starting from 65° C. to 55° C. in 10 cycles→1.5 min extensionat 72° C.)→25 cycles of amplification with a constant annealingtemperature at 55° C.→post-run extension (10 min at 72° C.). Foroxidase-domain amplification, the PCR program was the same except thatthe ramp temperature was from 60° C. to 50° C. and the extension timewas 45 sec. For PKS PCR, the extension time was 1 min instead. For PTSPCR, the program was identical to that for cyclase-domain amplification.For amplification of a large fragment containing both cyclase-domain andoxidase-domain at each end, CyFP and OxRP were used as PCR primer set.The ramp temperature was from 65° C. to 55° C., and the extension was 5min.

[0224] Subcloning and Sequencing

[0225] PCR mixtures were separated in agarose gels. Interested fragmentswere recovered by Gel Extraction kit (Qiagen, CA). PCR fragments weresubcloned into pGEM-T Easy vector (Promega, WI). Other restrictionenzyme-generated DNA fragments were subcloned into the appropriate sitesof pBSSK vector (Strategene), or pSP72, or pGEM-series (Promega).Recombinants plasmid DNAs were prepared by QiaPrep Spin Miniprep kit(Qiagen). DNA sequencing was performed by Davis DNA Sequencing. Inc.(Davis, CA). DNA sequences were analyzed with GCG package (GCG Inc. WI)and blasted against NCBI databases.

[0226] Library Screening.

[0227] PCR-amplified 1.1-kb cyclase-domain fragment was labeled withdigoxigenin (BMB) and used as probe to screen the cosmid libraryfollowing the standard colony-hybridization procedure (Sambrook et al.,supra). Positive clones were isolated and their cosmid DNAs wereprepared by alkali lysis method (Id). Subclonings from the left end ofCosmid 11 and the right end of Cosmid 1 (pYC9-2.1 and pYC25,respectively) were used as probes to perform cosmid walkings.

[0228] Cosmid Clone Mapping and Shotgun Subcloning.

[0229] Cosmid DNA was digested with BamHI, NcoI or AatII and separatedin 0.8% agarose gels. The patterns of fragments were analyzed togenerate a contig map. Southern hybridizations were performed to confirmthe validation of the initial screening and the mapping. All 18 BamHIfragments (except the liberated 8 kb cosmid vector) from two overlappingcosmids (Cosmid 1 and 11) were subcloned into the BamHI site of pBSSK)by shotgun cloning method.

[0230] Fermentation, Purification and HPLC Analysis of Leinamycin

[0231] Fermentation method was adopted from Hara et al. (1989) J.Antibiot. 42: 1768-1774. Fifty ml of seed culture was allowed to growfor 48 hrs at 22° C. with adequate shaking and used 10 ml to inoculate2×50 ml of fermentation broth. Culture was allowed to ferment for 48 hrsat 22° C. Moisturized Diaion HP-20 resin (SUPELCO, PA) was added to 5%(W/V) at 18 hrs after inoculation. Fermented broth was pooled and the pHwas adjusted to pH 2.0 with H₂SO₄. Resin was recovered from the broth byfiltration through two layers of cheesecloth. Crude Lnm preparation waseluted from the resin with 10 vol of methanol and solvent wasconcentrated by vacuum evaporation. HPLC separation of leinamycin fromcontaminants was performed with a Microsorb MV C18 reverse-phase column(Varian, CA) on a Dynamax SD-200 HPLC system (Rainin, CA). Authenticleinamycin (10 mg/ml) was provided by Tokyo Research Laboratories (KyowaHakko Kogyo Co., Japan).

[0232] PEG-Mediated Protoplast Transformation of S. atroolivaceus.

[0233] A 1.1-kb PCR amplified Cy-domain fragment was subcloned into: (1)pOJ260/EcoRI site to make the construct pYC12. (2) pKC1139/EcoRI site tomake the second construct pYC20; DNA of pYC12 and pYC20 was preparedfrom the non-methylated host strain ET 12567, denatured with alkalisolution and used for PEG-mediated protoplast transformation (Liu andShen (2000) Antimicrob Agents Chemother. 44: 382-392).

[0234] Conjugation between S. atroolivaceus and E. coli S17-1

[0235] Experiments of conjugation between S. atroolivaceus and E. coliS17-1 were performed by previous described procedure (Liu and Shen(2000) Antimicrob Agents Chemother. 44: 382-392) with modifications.Three exconjugates (named YC12C1, YC12C2 and YC12C3) obtained with pYC12were further purified, and the genomic DNA was prepared for Southernanalysis.

[0236]S. atroolivaceus cosmid Library Construction

[0237] High molecular-weight genomic DNA was isolated from S.atroolivaceus by a modified procedure (Rao et al. (1987) Methods inEnzymol. 153: 166-198; Kieser et al. (2000) Practical Streptomycesgenetics. The John Innes Foundation, Norwich, England). The majormodification was to extend the lysis time to up to 60 min. The cell wallof S. atroolivaceus seemed to be quite resistant to lysozyme treatment.Initial attempts that used a 30-min lysis time only obtained DNA sampleswith an average of ˜40 kb in size. Partial digestion by MboI was donewith decreasing amount enzyme. A portion of the digested samples wereanalyzed on agarose gel to obtain the correct size of genomic DNAfragment for library construction.

[0238] The S. atroolivaceus primary cosmid library contains about1.675×10⁵ colony-formatting-unit (cfu) in 500 μl SM buffer (335 cfu/μl),which is equivalent to 50 fold of the genome size. Analyses by alkalimini-preparation and BamHI-digestion of 16 randomly selected cosmidclones indicated that the average size of inserts is about 40 kb. Thelibrary was amplified once in XLI-Blue cells. Sterile glycerol was addedto the cell culture to 20% final concentration. Aliquots (10 ml each) ofthe amplified library were stored at ˜80° C. The titer of this amplifiedlibrary was about 10⁵ cfu/μl.

[0239] PCR Amplification of a Cyclase (Cy)-Domain.

[0240] An expected 1.1 kb fragment was amplified with the Cyclase-domainPCR primer set (CyFP+CyRP). Although the control reactions with singleprimer also yield amplified bands, the 1.1-kb fragment seems to be aunique band to the reaction with the presence of both primers. Thisfragment was cloned into pGEM-T Easy vector (Promega) as pYC1. Both endsof pYC1 were sequenced. Blast results indicated its homology to thecyclase-domains of known NRPSs.

[0241] Mapping the Lnm Cluster, Subcloning and Partial Sequencing

[0242] Digoxigenin labelled 1.1-kb Cy-domain probe was used to screenabout 4×1000 cfu of cosmid library, 15 well-isolated positive cloneswere obtained. Cosmid DNA from 12 clones was prepared and subjected toenzyme (BamHI) digestion, Southern hybridization and PCR diagnosis forthe presence of Cy-domain, Ox-domain, PKS-domain or/and PTS-domain.

[0243] Initially, four cosmids (#1, #2, #6 and #11) were found to coverthe longest chromosome region (total of 63 kb) and the PCR diagnosisdata supports the predicted domain-organization. Later, the other endsof cosmid #1 and #11 were subdloned as pYC25 and pYC9-2.1, respectively,and were used as probes to obtain more cosmid clones extending to bothdirections. The relative position of those cosmids has been mapped andis illustrated in FIG. 2. Subsequently, the sequence of the entire lnmgene cluster and its flanking regions (˜135 kb ) was determined andanalyzed (see FIG. 3). Deduced functions of the open reading frames(ORFs) in the leinamycin biosynthetic gene cluster are summarized hereinin Table 1 and Table 2.

[0244] The Development of a Genetic System for S. atroolivaceus

[0245]S. atroolivaceus grows very well at 30° C.; it doesn't grow attemperature beyond 35° C. It is found to be highly sensitive to bothapromycin (Am) and thiostrepton (Thio) (complete inhibition of growth at10 μg/ml and 3 μg/ml, respectively). Thus, E. coli—Streplomyces shuttlevectors pOJ260 (suicide vector, Am^(R)) and pKC1139 (self-replicatingvector, Am^(R)) were chosen to make the gene-disruption constructs pYC12and pYC20, respectively. The concentration of antibiotic (Am) forselection of transformants/exconjugates is 50 μg/ml.

[0246] TSB medium gives the best yield for protoplast preparation,however the regeneration ratio is very low (less than 0.01%).Protoplasts made from TSB with 30% sucrose has a regeneration ratio of0.2%. YEME gave the highest protoplast regeneration ratio of 0.6%. Allthe protoplast samples were generated on R2YE plates with 10 mM MgCl₂supplemented. Approximately 1×10⁹ protoplasts and 2 μg DNA were used foreach transformation experiment. Trial experiments were performed withpYC20 construct. Seven apparent positive transformants were obtainedfrom the initial selection plates. Only 1 remained well-growing after 2rounds of re-inoculation on vegetative-growth medium (TSB) andspore-generation medium (ISP-4). The calculated transformationefficiency was about 8×10⁻⁸.

[0247] Since the combined efficiency of transformation and regenerationwas only about 1×10⁻⁹, which is very low, a conjugation approach waspursued, with surprising success. Spores (1×10⁹) were heat-shocked for20 min at 42° C. instead of 10 min at 50° C., followed by incubation at30° C. for up to 6 hours. The germination of spores was monitored bymicroscopic checks every 30 min from 4 hours after heat-shock. E. coliS17-1 (bearing either pYC12 or pYC20) culture was freshly prepared.Conjugation was conducted on modified ISP-4 medium. After incubation at28° C. for 5 days, about 18 and 500 apparent positive ex-conjugates grewout from the initial selection plates with pYC12 and pYC20,respectively. Therefore, the calculated conjugation/integrationefficiency for non-self-replicating construct pYC12 was approximately1.8×10⁻⁸, and the conjugation efficiency for self-replicating constructpYC20 was 5×10⁻⁷. Three exconjugates (named YC12C1, YC12C2 and YC12C3)obtained with pYC12 were further purified and analyzed.

[0248] Genomic DNA hybridization confirmed the correct disruption of thetarget NRPS-gene gene in S. alroolivaceus. With the Cy-domain as probe,wild type gave a hybridized band of 4.6 kb in size with NcoI digestion,while all three independent exconjugates (YC12C1 to C3, appeared to beidentical) gave a band of 9.2 kb, which is the expected size aftervector single-crossover event. With the vector (pOJ260) DNA as probe,wild type DNA was not hybridized, while three exconjugates had the same9.2 kb positive band.

[0249] Analysis of Lnm Production in S. atroolivaceus Wild Type andMutants

[0250] Crude Lnm sample was extracted from 50 ml of 4-day 2-stepfermentation broth, following the described procedures (Hara et al.(1989) J. Antibiot. 42: 1768-1774). Lnm production was further analyzedby HPLC (FIG. 10). Samples purified from HPLC were also subjected toMass Spectrometry analysis. Lnm has the molecular formula ofC₂₂H₂₆N₂O₆S₃ and a molecular weight of 511.2 (Hara et al. (1989) J.Antibiot. 42: 1768-1774). The dominant MS peak was found in wild typeLnm sample but not in YC12C1. This confirmed the HPLC results that themissing HPLC peak is indeed that of Lnm (FIG. 10).

[0251] In summary, a 180 kb gene cluster encoding lnm biosynthesis wascloned from S. atroolivaceus, 135 kb of which was sequenced. Sequenceanalysis revealed that Lnm is biosynthesized by a hybrid nonribosomalpeptide synthetase and polyketide synthase system. These genes can nowbe used to improve the production of leinamycin, to engineer microbialstrains for the production of novel leinamycin analogs as drug leads,and to use as genetic materials in combination with other nonribosomalpeptide synthetase genes, polyketide synthase genes, and genes encodingother enzymes responsible for natural product biosynthesis incombinatorial biosynthesis to generate chemical structural diversity.

[0252] An efficient genetic system for in vivo manipulation of naturalproduct biosynthesis in S. atroolivaceus has been developed. Conditionsfor the introduction of plasmid DNA into S. atroolivaceus have beenoptimized for both protoplast-mediated transformation and E. coli-S.atroolivaceus conjugation. Genetic engineering of leinamycinbiosyntheses in S. atroolivaceus has been demonstrated by disrupting theNRPS module, resulting to the isolation of Lnm no-producing S.atroolivaceus mutants. The latter not only confirmed the cloned genecluster encoding Lnm biosynthesis but also demonstrated the feasibilityto making novel Lnm analogs in S. atroolivaceus by manipulating genesgoverning Lnm biosynthesis.

Example 2 A Discrete Acyltransferase Associated with a Type I PolyketideSynthase

[0253] In this example, we describe the in vitro loading of ACP domainsof leinamycin PKS by LnmG, a discrete AT. We have previously cloned thelnmGHI genes, demonstrated that this locus is essential for leinamycinproduction, and localized the lnm biosynthesis gene cluster to a172-kilobase (kb) DNA region from S. atroolivaceus S-140. DNA sequenceanalysis of the four overlapping cosmids, pBS3004, pBS3005, pBS3006, andpBS3007, revealed 72 open reading frames (ORFs). Sequential inactivationof ORFs from both ends of the sequenced region led to the assignment ofthe lnm gene cluster to consist of 27 ORFs, of which two (lnmQ and lnmP)encode nonribosomal peptide synthetase (NRPS), one (lnmI) encodes ahybrid NRPS-PKS, and one (lnmJ) encode a PKS (FIG. 11).

[0254] Materials and Methods

[0255] Sequence Analysis of the lnm Gene Cluster and Determination ofthe lnm Gene Cluster Boundaries.

[0256] The lnm gene cluster was previously identified and mapped to fiveoverlapping cosmids, pBS3004, pBS3005, pBS3006, pBS3007, and pBS3008.DNA sequencing of the first four cosmids yielded a 135,638 base pair(bp) contiguous DNA sequence. Bioinformatic analyses of DNA sequencewere done with the Genetics Computer Group (GCG) program (Madison,Wis.), revealing 72 orfs. The overall GC content of the sequenced regionis 72.4%. Functional assignments were made by comparison of the deducedgene products with proteins of known functions in the database andsummarized in the GenBank entry under accession number AF484556. Theboundaries of lnm gene cluster were identified by gene replacement oforf(−13), orf(−11), orf(−2), orf(−1), and lnmA for upstream boundary andof orfZ′, orf(+1), orf(+2), orf(+3), orf(+4), and orf(+6) for downstreamboundary, respectively. Inactivation of genes within the lnm genecluster abolished LNM production, whereas that of genes outside the lnmgene cluster had no effect on LNM production. LNM production andisolation from both the wild-type and recombinant S. atroolivaceusstrains were carried out as reported previously. HPLC analysis werecarried out on a Microsorb-MV C-18 column (5μ, 100 Å, 250×4.6 mm,Varian, Walnut Creek, Calif.), eluted with a gradient from 100% buffer A(20% CH₃CN, pH 3.6 with HOAc) to 68% buffer B (80% CH₃CN, pH 3.6 withHOAc) in 40 min at a flow rate of 1 ml/min and UV detection at 320 nm.

[0257] LnmQ and LnmP are NRPS adenylation (A) enzyme and peptidylcarrier protein (PCP), respectively, constituting the loading module;LnmI contains the previously characterized thiazole-forming NRPS moduleas well as PKS module 3 and the KS domain of PKS module 4; and LnmJharbors PKS modules 4 to 8 plus a TE domain. The Lnmmegasynthetase-templated synthesis of 1 (FIG. 11) could be envisaged tobegin at LnmQ and end with the cyclization of the full-length linearpeptide-polyketide intermediate by the TE domain of LnmJ to yield amacrolactam intermediate such as 2 (FIG. 11). Although it remainsunclear what the origin of the 1,3-dioxo-1,2-dithiolane is and how it isspiro-fused to the 18-membered macrolactam ring, subsequent modificationof 2 by the action of post-PKS enzymes could be envisaged to furnish 1(FIG. 11). The deduced Lnm NRPS and PKS functions are consistent withwhat would be required for the biosynthesis of 1 from the amino acid andacyl CoA precursors. However, the Lnm hybrid NRPS-PKS megasynthase ischaracterized by several intriguing features and, most strikingly, itlacks the cognate AT domain from all six PKS modules (FIG. 11).

[0258] Unusual Features Predicted on DNA Sequence for the Lnm HybridNRPS-PKS Melasynthetase.

[0259] The most striking feature of the Lnm hybrid NRPS-PKSmegasynthetase is the lack of cognate AT domain from all six PKSmodules. Others unusual features include: (1) a loading NRPS moduleconsisting of discrete A and PCP proteins, (2) tandem Cy domains for thethiazole-forming NRPS module, (3) tandem KS domains and the absence of adehydratase (DH) domain for PKS module 3, (4) tandem ACP domains and anintegrated methyl transferase (MT) domain for PKS module 6, (5) theabsence of a DH domain for PKS module 6 and of a DH and an enoylreductase (ER) domains for PKS module 7, and (6) the presence of anextra domain unknown to PKS between PKS module 8 and TE.

[0260] A PKS module cannot be functional unless its ACP domain is loadedwith the extender unit, and this step is catalyzed by the cognate ATdomain. Lack of AT domain in all modules of a type I PKS isunprecedented, raising the question of how the LnmIJ PKS modules arecharged with the extender unit malonyl CoA for the biosynthesis of 1(FIG. 11). Previously, we have demonstrated that lnmI is essential for 1production, inactivation of which abolished 1 production (FIG. 12C). Wesimilarly confirmed that lnmJ is essential for 1 production by replacinglnmJ with a mutant copy in which domains of PKS modules 7 and 8 weresubstituted with the apramycin resistance gene, aac3(IV). The resultantS. atroolovaceus SB3002 mutant strain lost its ability to produce 1(FIG. 22D).

[0261] To inactivate lnmJ, a 4,571-bp EcoRI-NotI internal fragment oflnmJ that harbors the KR and ACP domain of PKS module 7 and the KSdomain of PKS module 8 was replaced with the aac(3)IV apramycinresistance gene and cloned into pSET151 (Bierman et al. Gene 116;43(1992)) to yield pBS3019. The latter was introduced into S.atroolivaceous S-140 by conjugation and selected for apramycinresistance and thiostrepton sensitive phenotype to isolate the doublecrossover mutant strain SB3002, whose genotype was confirmed by Southernanalysis. To inactivate lnmG, a 388-bp SalI-BamHI internal fragment oflnmG was replaced with the aac(3)IV apramycin resistance gene and clonedinto pSET151 to yield pBS3020. The latter was similarly introduced intoS. atroolivaceous S-140 to isolate the mutant strain SB3003, whosegenotype was confirmed by Southern analysis.

[0262] To search for the missing AT activity, we re-examined geneswithin the lnm cluster and identified lnmG, the deduced product (theN-terminal half) of which is highly homologous to AT domains. Weinactivated lnmG by replacing it with a mutant copy in which lnmG wasdisrupted by aac3(IV). The resultant S. atroolivaceus SB3003 mutant lostits ability to produce 1, confirming that lnmG is essential for 1production (FIG. 12E). These results suggested that LnmG provides the ATactivity in trans to LnmIJ and loads the malonyl CoA extender unit toall ACP domains of the six PKS modules for 1 biosynthesis.

[0263] Expression of lnm Genes in E. coli and Purification of theResultant Recombinant Proteins.

[0264] To validate the activity of LnmG, we expressed both lnmG and theseven ACP domains from the six PKS modules encoded by lnmIJ as well aslnmP in E. coli and purified the resultant LnmG (FIG. 13A), ACPs andLnmP PCP (FIG. 13B) as His₆-tagged fusion proteins to homogeneity. (Weincluded the LnmP PCP as a negative control to demonstrate that LnmGdiscriminates the LnmIJ PKS ACPs from other carrier proteins.)

[0265] The lnmG gene, the seven ACP domains from lnmI and lnmJ, thetridomain PKS module 4 of lnmJ-(DH-ACP-KR), and the lnmP gene were allamplified by PCR with primers listed in Table 3 and cloned asNdeI-HindIII fragments into the same sites of pET28a (Novagen, Madison,Wis.), yielding expression constructs pBS3021 to pBS3030, respectively(Table 3). Sequence fidelity of PCR products was confirmed by DNAsequencing. Introduction of pBS3021 to pBS3030 into E. coli BL-21 (DE-3)resulted in the overproduction of these gene products as His₆-taggedfusion proteins, respectively. The latter were purified by affinitychromatography on Ni-NTA resin under the standard conditions recommendedby Qiagen (Valencia, Calif.) and subsequently dialyzed against 25 mMTris-HCl, pH 7.0 (for LnmG) or pH 8.0 (for ACPs or LnmP), 25 mM NaCl,10% glycerol, and 2 mM DTT and stored at −80° C. TABLE 3 Summary of Inmgene expression constructs Gene¹ Primers² Construct InmG Forward:5′>CGGAATTC CAT ATG GTG GCA CTG GTT TTC CCG>3′ pBS3021 Reverse: 5′>CGGCCAAG CTT CCC CCC GGC GAG GAC GTC>3′ InmP Forward: 5′>CGGAATTC CAT ATG TGGGAC CAC AAG TTC GAG >3′ pBS3022 Reverse: 5′>CGCGC AAG CTT TCG GCC GGCTCC GTC GAG >3′ InmI-ACP3 Forward: 5′>CGGAATTC CAT ATG TCA GTC ACC GGGCCG CCC >3′ pBS3023 Reverse: 5′>CGCGC AAG CTT CCC GAG GTC CGC CAGATG >3′ InmJ-ACP4 Forward: 5′>CGGAATTC CAT ATG CCC CCG GAC GCG GTGCGC >3′ pBS3024 Reverse: 5′>CGCGC AAG CTT GAA CTC CCC GTA CAG GTG >3′InmJ-ACP5 Forward: 5′>CGGAATTC CAT ATG GAC CCC CAG GAG GTC CTG >3′pBS3025 Reverse: 5′>CGCGC AAG CTT GTG CAG TTC CCT GAC GTG >3′InmJ-ACP6-1 Forward: 5′>CGGAATTC CAT ATG TCG CCC GAG GCC GTG CCC >3′pBS3026 Reverse: 5′>CGCGC AAG CTT GTG TTC CTG CCC GAA GTA CC >3′InmJ-ACP6-2 Forward: 5′>CGGAATTC CAT ATG TCC CCC GAG TCC CTG CCC >3′pBS3027 Reverse: 5′>CGCGC AAG CTT GTG CTC GGC GCT CAG GTA C >3′InmJ-ACP7 Forward: 5′>CGGAATTC CAT ATG CTG CCC GAG CTC GTG GAG >3′pBS3028 Reverse: 5′>CGCGC AAG CTT ATG GTG CTG CGT CAG CTA CT >3′InmJ-ACP8 Forward: 5′>CGGAATTC CAT ATG GCC GCC TCC ACC GTC GTC >3′pBS3029 Reverse: 5′>CGCGC AAG CTT CAC CAC CCC CCC GAC CAA C >3′InmJ-(DH-ACP4-KR) Forward: 5′>ATGAATT CAT ATG AAC CTC CCC TCC CCA C >3′pBS3030 Reverse: 5′>AT AAG CTT GCC GTC CGG GGA GTC AGG >3′

[0266] Since most ACPs or PCPs overproduced in E. coli are in thenonfunctional apo-forms, we incubated them with CoA and the Svpphosphopantetheinyl transferase to ensure that all carrier proteins areconverted into the functional holo-forms (Walsh et al. Curr. Opin. Chem.Biol. 1:309 (1997); Sanchez et al. Chem. Biol. 8:725 (2001)). Weincubated the holo-carrier proteins with [2-¹⁴C]malonyl CoA and LnmG todirectly test malonyl CoA extender unit loading. The reaction mixtureswere subjected to SDS-polyacrylamide gel electrophoresis and phosphorimaging to detect specific loading of the [2-¹⁴C]malonyl group to thephosphopantetheinyl group of ACPs (FIGS. 13C and 13D).

[0267] In vitro Assay of LnmG with ACPs or PCP and [2-¹⁴C]malonyl CoA

[0268] LnmG-catalyzed loading of the malonyl group from malonyl CoA toACPs or PCP was assayed in a two-step reaction. First, thephosphopantetheinylation of apo-ACPs or PCP was catalyzed by Svp in thepresence of CoA (Sanchez et al. Chem. Biol. 8:725 (2001)). A typicalreaction of 75 μl contained 100 mM Tris-HCl, pH 7.5, 12.5 mM MgCl₂, 2.5mM DTT, 33.3 μM CoA, 10 μM ACP or PCP, and 2 μM Svp and incubated at 25°C. for 60 min. Second, a mixture of 2 μM LnmG and 10 μl of[2-¹⁴C]malonyl CoA (200 μM, 51 mCi/mmol, PerkinElmer, Boston, Mass.) in15 μl volume was added to each reaction. Reaction was incubated at 25°C. and subsequently quenched by addition of 900 μl acetone at varioustime points. Proteins were precipitated by centrifugation at 4° C. for30 min after frozen at −80° C. for at least 1 hour. Protein pellet wasre-dissolved in 1×sampler buffer and separated on 4-15% SDS-PAGE gels(Bio-Rad, Richmond, Calif.). The resolved gels were visualized byCoomassie blue staining and phosphor imaging (LE phosphor screen,Amersham Pharmacia, Piscataway, N.J.).

[0269] Alternatively, the reaction mixtures were subjected to highperformance liquid chromatography species were purified and subjected toelectronspray ionization-mass spectrometry (ESI-MS) analysis (Table 4).For HPLC preparation of apo-, holo- and malonyl-ACPs, thephosphopantetheinylation reaction or complete loading reaction wasscaled up 3 times and cold malonyl CoA was used instead. The second stepof loading the malonyl group to holo-ACP was proceeded for 10 min at 25°C. HPLC analysis were carried out on a Jupiter C-18 column (5μ, 300 Å,250×4.6 mm, Phenomonex, Torrance, Calif.), eluted with a gradient from85% buffer A (H₂O+0.1% TFA) to 90% buffer B (acetonitrile+0.1% TFA) in25 min at a flow rate of 1 ml/min and UV detection at 220 nm to separatethe proteins in the reaction mixture. Individual protein peaks werecollected manually, lyophilized and re-dissolved in H₂O for MS analysis.ESI-MS analyses were performed on an Agilent 1000 HPLC-MSD SL instrument(Palo Alto, Calif.).

[0270] LnmG specifically and efficiently catalyzes the loading of themalonyl CoA extender unit to the LnmIJ PKS ACPs, and no loading wasobserved in the absence of LnmG. After 5 minute incubation in thepresence of LnmG and [2-¹⁴C]malonyl CoA, six of the seven ACPs wereefficiently loaded with the malonyl group (FIG. 13C, lanes 2-4 and 6-8)with ACP6-1 being loaded less efficiently (FIG. 13C, lane 5), and noloading was observed for the LnmP PCP (FIG. 13C, lane 9). [Theobservation that ACP6-1 is less efficiently loaded is consistent withthe finding that PKS module-6 has two ACP domains, one of which may bethe preferred site for malonyl CoA loading (FIG. 11).] LnmG was alsolabeled by [2-¹⁴C]malonyl CoA (FIGS. 13C, 13D, and 13G).Acyltransferases are known to form acyl-O-enzyme intermediates at theiractive site Ser residues before transferring the acyl groups from theirCoA substrates to the nucleophilic recipients such as an ACP, andlabeling of acyltransferases by extender units has been confirmed forboth the AT domain of type I PKS (Marsden et al. Science 263:378 (1994))and the malonyl COA:ACP transacylase of type II PKS (Carreras et al.Biochemistry 37:2084 (1998)). Interestingly, the extent of ACP labelingdecreased with longer incubation time—reaching a maximum in the first 5minutes and falling to less than 10% after about 2 hours as exemplifiedby LnmI ACP3 (FIG. 13D). The malonyl-S-ACP product is apparently notstable under the assay condition, the malonyl group of which undergoeshydrolysis in the absence of chain elongation. Analogous results havebeen found for both the DEBS PKS (Marsden et al. (1994)) and fatty acidsynthase (Wakli, S. J. Biochemistry 28:4523 (1989)). In contrast, LnmGlabeling appeared to be constant (FIG. 13D), suggesting that themalonyl-O-LnmG species is stable under the assay condition. To excludeany ambiguity associated with these assays, we subsequently purified theapo-, holo-, and malonyl-S-ACP species from the assay mixtures by HPLC,as exemplified by the LnmI-ACP3 (FIG. 13E), and established theiridentities by ESI-MS analysis. As summarized in Table 4, distinct [M+H]⁺ions at m/z values exact or near the calculated values of thecorresponding ACP species were observed for all samples analyzed,confirming their predicted molecular structures. TABLE 4 ESI-MS analysisof apo-, halo-, and malonyl-S-ACPs apo-ACP holo-ACP malonyl-S-ACP [M +H]⁺ [M + H]⁺ [M + H]⁺ ACPs¹ Calcd. Found Calcd. Found Calcd. FoundLnmI-ACP3 11,702 11,700 12,042 12,040 12,128 12,126 LnmJ-ACP4 12,24512,241 12,585 12,582 12,671 12,669 LnmJ-ACP5 12,520 12,517 12,860 12,85712,946 12,943 LnmJ-ACP6-1 12,209 12,206 12,549 12,546 12,635 12,632LnmJ-ACP6-2 12,151 12,147 12,491 12,486 12,577 12,572 LnmJ-ACP-7 12,32212,318 12,662 12,665 12,748 12,751 LnmJ-ACP-8 12,090 12,087 12,43012,427 12,516 12,512

[0271] Finally, we expressed the PKS module 4 as a tridomain protein,LnmJ-(DH-ACP-KR), in E. coli and purified it as a His₆-tagged fusionprotein (FIG. 13F). LnmJ-(DH-ACP-KR) was similarly assayed in thepresence of LnmG and [2-¹⁴C]malonyl CoA to confirm that LnmG can loadthe malonyl group to a multi-domain PKS module as efficiently as toindividual ACP domains. As summarized in FIG. 13G, LnmG efficientlyloaded the malonyl group to holo-LnmJ-(DH-ACP-KR) (lanes 3-6); theextend of [2-¹⁴C]malonyl labeling reached maximum in 5 min (lane 4) anddecreased with longer incubation time as a result of hydrolysis (lanes4-6), and no loading was observed with the apo-LnmJ-(DH-ACP-KR) protein(lane 2).

[0272] Yield Improvement by Overexpression of LnmG

[0273] Given the mechanism that LnmG is responsible for the loading ofthe malonyl CoA extender unit to all six PKS modules of the Lnm NRPS-PKSmegasynthetase, we reasoned that LnmG could be a rate-limiting factorfor 1 biosynthesis. We therefore explored yield improvement for 1 byoverexpressing lnmG under the constitutive ErmE* promoter in both low-and high-copy-number vectors in S. atroolivaceus SB3003 (Kieser et al.,Practical Streptomyces Genetics (The John Innes Foundation) Horwich2000). To construct the lnmG overexpression constructs, a 450-bpEcoRI-SacI fragment that harbors the ErmE* fragment (Ribb et al.,Mol.Microbial. 14:533 (1994)) and a 2,883-bp SacI-BglII fragment of thelnmG gene were cloned into pBS3031 [a low-copy-number vector derivedfrom the SCP2* origin of replicon (Kieser et al. (2000)) and pBS3032 [ahigh-copy-number vector derived from the pIJ101] vectors, respectively,to yield pBS3017 and pBS3018. The latter were introduced into S.atroolivaceus SB3003 by conjugation, and the resultant SB3004 and SB3005strains that harbor pBS3017 and pBS3018, respectively, were cultured andanalyzed by HPLC for LNM production as described previously with the S.atroolivaceus S-140 wild-type strain as a control). The resultantrecombinant strains SB3004, harboring the low-copy-number expressionconstruct pBS3017, and SB3005, harboring the high-copy-number expressionconstruct pBS3018, produce 3-5 folds more 1 (FIGS. 12F and 12G) than thewild-type S-140 strain (FIG. 12B) as determined by HPLC analysis.

[0274] It is understood that this invention is not limited to theparticular methodology, protocols, cell lines, vectors, and reagentsdescribed, as these may vary. It is also to be understood that theterminology used herein is for the purpose of describing particularembodiments only, and is not intended to limit the scope of the presentinvention which will be limited only by the appended claims.

[0275] It must be noted that as used herein and in the appended claims,the singular forms “a”, “an”, and “the” include plural reference unlessthe context clearly dictates otherwise. Thus, for example, reference to“a cell” includes a plurality of such cells, reference to the “vector”is a reference to one or more vectors and equivalents thereof known tothose skilled in the art, and so forth.

[0276] Unless defined otherwise, all technical and scientific terms usedherein have the same meanings as commonly understood by one of ordinaryskill in the art to which this invention belongs. Although any methodsand materials similar or equivalent to those described herein can beused in the practice or testing of the present invention. All patents,patent applications and publications mentioned herein are incorporatedherein by reference for the purpose of describing and disclosing thepolypeptides, polynucleotides, cell lines, vectors, and methodologieswhich are reported in the publications which might be used in connectionwith the invention. Nothing herein is to be construed as an admissionthat the invention is not entitled to antedate such disclosure by virtueof prior invention.

1 214 1 135638 DNA Streptomyces atroolivaceus 1 cgaagcggaa gcgcccaggtcagtctggct gtagcggcgg cccaggtgaa cagtgcatct 60 cgactgcgac gagccgtcattgaccgtggt tgaccgcacg atgtggcacg gctgtggcac 120 acgaccgctt cgtcccgaagcaactcgggg cgctcgctgt ccttgggctg ctgcgcctgt 180 caccggcgct gatggtccgctgacgtccac ggttgttcgt ggttgtgcgt cgcggttgtc 240 acgcagctgg acactcaccactactactgt ccgtgtgagg cccgaaaggt tctcgtaaat 300 gatcttcgac cggtgtcgaccgcctagtgc gaggcgaaga tcgcctgatg cccgatcgtg 360 atccaagggt gtacaaccccgtgcggatcc caactgtctg aagggcgccg gaggatgagc 420 ctccggcgct cttccgtacggagatcgggg atcaggtggg gaagaagatc tggagagctg 480 tcgcggccgg tggcgcggcggcggtgttca cggccatggc cgcgggaccg gccggggcgg 540 ccgagggcgg ggtctcgttcacccgtgtcc aggtgaacgg cggcaaggcg atcgtgattg 600 gggtgtcgaa cgaggtcgcggtgcccacat ccttccggat ggcgaccaca caccagtgga 660 agtggccggc ggtgttcctgtaccgcggca agacgagtga gcggctgtgg cacgcgatcg 720 agacgagcga ctgcatcaaggtgagctcgg gcgtctgtga cttcaacgag acgatgtact 780 tcgacccgag cgagtgggagatgcgcaaca gcgaggccgg ggcgtggaag gtcgcggccg 840 aggtctactt ccagggcggcggcggcgaca ccgatgacga gggcctcacg gcctacgtca 900 agcgcaactc ccacctgaccgtgaacgcct cgcccgaacc ggtggccaag gacaagacca 960 tcaccgtgac cggcaaggtcacgcgggcca actgggagac caggaagtac gtctcctacg 1020 aaggccgcct ggtgaccctgcagttcaagc ccaccggaac cacctcctac accacggtga 1080 agaaggtgta cgcgaacggctcgggcaacc tcaagacgac cgtgaaggca tccaagacgg 1140 gcacatggcg ctgggtgtactacggcaaca ccactaccgg gccgtcgact tcggccggcg 1200 acaatgtcgt agtgaagtgagttccctgga ctgcctctcg acctactcgg cacagctaca 1260 ggacaacctt taggctcggcttcgcgcggt cggcggcgag gtatccagct ctgaagtgca 1320 gctgcatccc tgattctgtctaggggtggt ccggtatcca atccgtggcc caaccgacct 1380 ccaggcctag ttgatagctatgagggggac ctgtcagcgc cgaccccgca caagccctga 1440 acggtcttga aaaccgtcgtggcgcaagtc gctttgggtt caaatcccac acccaccgcc 1500 aggagaaacg gcccctgaccaggcattttc agtcaagggc cgttctgctg tccactcccc 1560 gccagtgact gccattccccgggtgatcgg gcacgagagg ggcacggcta ccttctgatc 1620 cgtagctcta tatagatcggtcagcgacag tcattccggt cgcttcacgg cccaggagag 1680 acgcaaccgg tcgggggcggccgatggggt tgaggtactt cgctgctgta cgccgccccg 1740 ggagacgacc ccgagtgcgagcaggtctac tctcctcgga tggacgacgc ccctttcttc 1800 gcctcctgcc gggccgtcgcacggctggca cccagctaac catccacacc accggagatc 1860 gtttgggtcc gctccacgaaaaggcggaag tcttccagag gtgcaggtcg cagtggggcg 1920 gtcacttcag gctgtaacgcgtgttcctgg tctcggaggg ggccagacta ggccgcacct 1980 ggtgtgcttg gcttgattaaagccacgaac gtaccccgct ttcataccgg cgagccaacc 2040 tttattgtag gcctcgaccctagcctggtc agtctcgttt tcattttcca gctgacgtcc 2100 gcacccggcg tcgaagccgcggctgaaacc gtgagcgtcg ccctcaaagt atccttcgtt 2160 gtagtcagtt atgtatttgttctcagttat gtatgggcac ttcgcaccag cgggggcagc 2220 gcaaggtctc gaccagggtgctgcggacgc ctgagtgatg acatgaggag ataaagcgag 2280 agtagccgca acgagcgctggactgagccg aatgaatagt cgtctggcat ccatggttct 2340 ccccttggac caactcggctttggggcggc gatcgtctgg ctgtcatcga cgccaggtcg 2400 ccgcctctaa tcgatcgcgaggttccacct ctactgctgt tcccatggtg cgcttcagcc 2460 ccaaggctgt catctcgtgttgaatcaacc gctgtgaggg ccgaaagcct ccgccgtccg 2520 cgcacgcata ccggcaagacgggtgattcg tgataccaca ccgcgccacg cccctcaacg 2580 cgcagtgccg gacgcacgagcccatcgcga ttcacaacaa atggcctatc agagtctcga 2640 agacgtccgg ccatcctgcggccgttcttg tatccgggca ctcactccga agccatggca 2700 aacgcccgga tcaagtgggaggccgtacac cactcctcag gacgcaaccc ggccttgaaa 2760 accgtcctgg cgcgagtcaccgtgggttca aatcccacac ccaccgcgca ggttgaaggc 2820 ccctgaccag caacattggtcgggggcctt cgctttacac gctgtccgtg agtgaccgtc 2880 gctccccggt gttccccgctcgatcgggca cgccgggggc acggtcagtc ttcgacggga 2940 ctgggcgccg tgaggtcaatccatatccac ggcttgtgat cccagtcgcg caccttgcct 3000 atgactaggt cgacggtcgcccccaccggc ggcagctcct gcacgagtcg cttcacgccc 3060 ggctcgcttc tgcggcgaatcgtgtccagc gaggcgccca cggggtcgta ctcgttgagc 3120 tttgccatga ggccccatggctcgtgacta gtgatcgttg cttggactgt ctcgccttgc 3180 tgaagctcgc tgagagggcgtctcggttta gacgctatct cccgttccgg gcggttcgcg 3240 gtccgattcg gagagtgcgtacaagaaggc aaatcccttg cccttaagtc gctcgcgtca 3300 cgttgccttg tgctgacactcttcgacctg gacaacaccc tgatcgaccg gcacaggcct 3360 tgagtactgg gctcgtgacttcgtccgctg gcgctctctg cctcaagagg cggaggcagc 3420 catccggccg ggcctccccgcagccccatg ctgcagacat ctgaagcgga aactgaggag 3480 accaccgaat ggagatgtccgacacctcgc gtgcgatcgc tgctgcgatt tcgattgccg 3540 catctctcgg cctgccggccgatgacgcga tcgttctcca caactcgaac aagctggcac 3600 tgcggctgac gccgagcgatgtccttgccc gggtcgcccc tgtgggacaa gaggttgcac 3660 acttcgaagt cgagctcgctcaaaggctcg ccgagttcgg atctccggtg ggcctcctgg 3720 aacctcgggt ggacccgcgtgtgtacacgc gcgacggctt cgccgtgacg ctgtggacct 3780 actacgagcc cgtgacacctcacgtctcac cagccgacta cgccgaggcg ctggagcgac 3840 tgcacgctgg catgcgcaaggtcgatgtgc cgagcccgag cttcatggag cggatcacgg 3900 aggcgcaaga agtcgttgccagcctggatc tctcgccggg gctagccgac gcagaccgtg 3960 tgctcctcag cggcaggctgggaagcctgc gacgggcgat cgagggccgc ggcgcggtgg 4020 agcagttgct ccacggcgagccgcatccgg gcaatgtgct cagcacgaag aagggcccgt 4080 tgttcatcga cctggagacgtgctgccgtg gtcccgtcga gttcgacctc gcccacgctc 4140 ccgaggaggt ctgcaagcactaccccaacg tcgaccaagg gctgctcgat gactgccgac 4200 agctcgtcct cgcgatgctcgcagcatggc gttgggagct cggcgaccag ttcccgaacg 4260 ggaagcgatt cggggaagaatttctccgct tgctgcgaga gggcccaccc tggccgacct 4320 tcgacgcagt gaccaggcgactgggcagcg gccagtagaa atcaccaatg tggcacgacc 4380 gggatcaccg ccatcgtccgagccgtcatc gttcttgaac atgccagctg ggcaagatgg 4440 aaaggcccag tggctcagacttccggcggg attgccagca tctgatccgt aggcgggtgc 4500 gcgcacggat cgcctgaactcccctagcca tcagggtctg ttggtcaggg gtcacctgta 4560 catcgacatg tgcatgtgtgcgccgacgtt ggtgtccgac agcggcgctt ccgaagggtc 4620 gggcacttga ggctgggccacgccccaccc ctcaacaggg tgacctggga aatcccttgg 4680 ctcggaccac cgcttcaggagttcgatccg aagggccacg ctgggccgat agccaacttg 4740 gcagccacgt cgaaggccgcggttggccac tgctgttcgt tggcgttgat gtcagacaca 4800 gatgtcggta agcctcactaccagcccccg tcgaaccgag gttgcagggt ggccgccacg 4860 cccgacgtga cagcgagtatgccgcccggg ccggccaggg tggtcgtctc gcgttccgga 4920 acaactcgtg ctggagcgtcggcgaagtcg atgcacgacg ccgggagctg cggcgtcagg 4980 caccttggcg gtggtgccgagcgtgtcgca agagacccct gcgatcaccc ttgctaccgt 5040 ccacctcgcc taaataatccacaattgacc gattcgcacc cgcgggtgtt gcgagaggac 5100 acggaccatg agcgacttttcccgccgccg actgctgaag acttccgcgg tcgccggcgc 5160 cggcgcagtc gtcgtgccgggcgtcgcggc agccgcgacc cctagtgcgg gtgtcgtgac 5220 ggaggcttcc gtaaatgaggcgtacgaggc gcagtgttcg ccggcgaagc tgaccggccg 5280 catcgtccgt ccccacgaccctgggtacac ggatgcgcgc ctcggctggg accagctctt 5340 ctctcactat ccgctggtcatcgtcttcgc tcagaacacc caggacgtgg tcaacgccct 5400 gacctgggcg cggcagaatgatgtcgcgct gcgaatccgg agcggccgcc acagcctaga 5460 gggctggtcg aacgtcgacagcggcctcgt gatcgacatc agcgaactga agtcggtcca 5520 tctcgacagc gctgctcgtgtcgcgacggt cggcgccgga ctcaaccagc tggaagcggt 5580 gaccacgctc gcgaagcggaacttcgcggt gacgacaggc accgaaggca cggtgggcct 5640 ggccggcgcg acgctcggtggcggcttcgg cttcctgacg cgctggctgg ggatggcgtg 5700 cgacagcctg atcggcgccgaggtcgtggt ggccgaaggc ggcgagtgcg caaaggtgat 5760 caaggccgat ctgacccataactcggactt gttgtgggcg ctgcgcggtg cgggcaacgg 5820 caacttcggg atcgtcacctcactcaccta caaggtggct ccactgaaga gcgttacgta 5880 tgtgcaggcc acgtgggatggcatcggtga cctgcggagg gtcttcaatg cctggcagcg 5940 caccgcgctg cacgccgatgaccgccttgg cacgcaggtc gaaatccacc gcaatcagac 6000 cttgttgttc gcggtcctcgcggaggggac acctgcggag gccaagaagc tactggcccc 6060 gatcctgtcg gtcggcagcccccaggtgtc ggtgcaggtt ggcaactggg gcgaggtgta 6120 ttcgggattc cagattcctaccgcgaacga acccgcgaac tggaagttct actcgcagtt 6180 caccaggaag ccgttcccgggcaaagcgat cgacgtgatc tgctcgttca tgaagcacgc 6240 gcccacggac gacagcaacttcttcactca ggcgttcggt ggggcggtca gaaggagtcc 6300 ccgcggcggc acggcgttcccgcatcgcga cgcactcttc tactccgaac ccggtgccgg 6360 ctggggaacc cgttccgaccagccgggcat ctgcgacccg ctcaccccgc aggcgcaagc 6420 ctggatcgcc gagttcagccaggccttgcg cccctacgtg gacggtgcct acgtcaacgt 6480 gccgaacgtc ggcatgcaggactgggaaac cgcctactgg aggggcaact tcgaccgcct 6540 gcgcgagatc aaggcgaagtacgacccgca caacgtcttc cagtacgacc agagcatccc 6600 gcctgcgtcc cgctgaccgaacttgatgga gccccctccc gaccggtgag ggggctccgc 6660 tgcgcctacc cgtctcaggcctttcgctca agagagccgg actttgatgc ggttcgacaa 6720 tccgtggccg gagcgcccctttctcgacga tcagccgatc ccgcacgcca tttggggact 6780 ggtcggagga gcccaccggctgctggccgg tccttggggt gcggatgtgg tgcgctctcc 6840 tttgcgcgct ggcgaccactgagccgtggc cctccgccgt acagacgggg tccggtgggg 6900 actactcagg acgccgatcgcgcgccacgc tcgtacggcc accccgcgag cgcctcttgc 6960 tcccaccggt tcggcttgcgcctcgggcgc ccccgggcac ctgttgtcgg gacggccgac 7020 tctatggccg cagggaactacggccaagcg cttaccagtg cggggggggc gagggcacgc 7080 gctgagtgac ggcgagtgtgtaccggccgt ggggctgtcg cccctgcggg aacgcccatc 7140 gcccatcgat gacgtgagagcgcgggcccg ccgcacctgg cgtccgcccg gcggaggcca 7200 tgcgccaagc cgccgaccgggatgccccac aggcccgttg cgccgtccct cagcacgccg 7260 aattcatggg cggtccgcccacccggacgc gcggtggccg acgcccccgt ctgctcgtgg 7320 cgtaactgtg acatggcacaagagcgacat gtgcgacccc ttgaacgacg gcgggggagg 7380 ccggaggatc cagtgatctttcgctcttcg ccgcgtctca ggagtcttcc ttgcccaagc 7440 ttcccatccc ccggcccgggccgatcgccc tcgccgtggc ggccgcgctg acactcggcg 7500 tcataccggc gactgccgctgtgtccgcgt actcgtccgg tgaggtgtcg cacaacgacg 7560 ccggacaccg aacccccggcgatgccggca cgcacaccgt caccctgatc acgggtgaca 7620 aggtcaccat cgggaccgccgccgacggca ccgtcgtgcg ctccttccgg gccgcgagcg 7680 gagccaccac cggcttccaccgcgctgtcg tggacggcgc cacctatgtc taccccgacg 7740 ccgccctccc gtacgtgagcgccggcaaac tcgacaagca gctcttcaac gtgacgcgcc 7800 tgatcaccga gggttacgacgacgcgcaca gctcccggct gccgctgatc gtgcgctaca 7860 ccggagccgc cgccaaggcccgcacccgga cgaagatggc cggctcgacc gacgtccgcc 7920 gcctggacag catccagggcgcggccctcg cgcagaacca caagcaggca ccggagttct 7980 ggtcctcgct caccgggggctccggcgcgg cggcccggtc cgcgaagccc tccttcgcgg 8040 gcggcgtcgc ccacgtgtggctcgacggca gggtgaaggc cgatctggcc gactccaccg 8100 cccagatcgg cgcgcagaaggtatgggcgg agggcgacac cggccaggac gtgaaggtcg 8160 cgatgctcga cagcggcgcggacaccgaac acccggacct ggtcgggcag gtgtccgaca 8220 gcgccagctt cgtccccggcgaggacgaca tcgccgacta caacggccac ggcacgcacg 8280 tcgcctcgac catcgtcggcacgggcagcg cctccgacgg caaggagcgg ggtgtcgcct 8340 ccggcgcccg gctgtccgtcggcaaggtgc tcaactccga gggcagcggc caggaatcgt 8400 ggatcatcgc gggcatggagtgggccgccc gcgaccagaa ggccaggatc atcagcatga 8460 gcctgggcgg cggcggtgacaagaacgacc cgatgagcca ggccgtcgac gaactcagcc 8520 acgacacggg cgcgttgttcgtgatcgcgg cgggcaacgg cggcccgcac tccatcagca 8580 gccccggtgc ggcagactccgcgctgaccg tcggcgccgt cgactccacc gacacgctcg 8640 ccgacttctc cagccagggcccgcgtgacg gcgacggcgg gctgaagccg gagatcaccg 8700 cgcccggcgt cgacatcgtcgcggcgcgct cgcactacaa gcgcggctcc ggctactaca 8760 ccacgatgag cggcacgtcgatggcgacgc cgcacgtcgc cggtgtcgcc gcgctcctcg 8820 ccgccgagca ccccgactggacgggcaccc agctcaagga ggcactggtc agcagcgcca 8880 aggcaacgcc ggcgtacaccccgtaccagg cgggcgccgg ccggctcgac gcgccggcgg 8940 ccgtgcacac cacggtcttcgccaccacga ccgcctactc cggcttccac acgtggcccc 9000 cgaagcccgg ggagaccgatgtccggacgg tgacgtacac caacgtcggc gacgccccgg 9060 tcagcctcaa cctggccgtcaacggcaccg tcccggccgg gttgttcagc ctctccgagg 9120 accatgtcac cgtgcccgcgcacggcaccg ccacggtcac cctgaccgcg gctctggaca 9180 agctggcggg cgaccagtcggtcagcgccg tgatcaccgg tacggacggc agcggagcgg 9240 tccgtgcccg gactctgatcggcgcggccc gagagggcca gcggcagaat ctgaccgtcg 9300 tcgcgaagga ccgtgcgggcaagcccctgg cgggccgggt cgtcctcacc gccgagcacc 9360 tcttcacggt gatggacctcgacgcgtccg gcaccggcac gacgcggctg cccgtcggca 9420 gctacagcgg ctggctcaccggggacgtgc agggcgccga aggcccgaac tcgctcggca 9480 tggtgctgct cgccttcaacgacgtgaagc tggaccagga ccgcaccgtc acgctggacg 9540 gccgcaaggc gcgccggatcctggcgcgcg tgccgcagca gaccaccgcc gtcgcgccgc 9600 gcctggacgc ccaccggtccttcaccgaca gcctggtgga gacgtcgatg ctgcccaact 9660 cgtcgtacga cagcatctgggcgctcccga ccggcaagaa ggtcaccgac ggcgagttcg 9720 aggccggcgc ccgcttccgcctcgagcagc cggcgctgac cttgaacacg aagtcgacga 9780 ccttcaacga cccgttggtcaagcgcgcgg ccacgccgct gcccgccggt agccggaacc 9840 tgacggcggt cttcgccggcgacggcacgg ccgaggagct cgcgcggcag aacgtgcgcg 9900 gcaaggccgt cgtggtgcggggcgccggca cggaagggat caaggcgcag gcggaggccg 9960 ccgcggcggc cggcgcccggctgctgctgg tcgtcaacga cggcgtcggc cgcctgcagc 10020 cctgggtcga gaacccctggagcccgcaga gcccggcccc ggtgacggtg gcgacgctca 10080 ccgccgacca gggcgcggagctgatcggct cgctccggca cggcgcggtg tcgctgaagg 10140 tcacctcgca ccccacgacggactacgtct acgacgtggt gcaccactgg accggcgccg 10200 tcccggcgga cccgacctggcgtgaggagt ccaaggacct cggccgggtg aacgtctcct 10260 tccggaacta ccgtcagggcaaggcgatgg agttccgtcc ggacgtctgg cgcggctggg 10320 ccgtcggtaa ccagctcacggcgcccgccc agggcgagcg caccgactgg gtcacctccg 10380 gcaccacgtg gctggacgacgccttcatcc agggcgagac cgggcagcac tcgatcgacc 10440 cgctgcagta ctcggcgcgcaagaccggca aggtcaactg gttcggcccg atccagcggc 10500 cccgcatggg cccggtgaactaccagcccg tgcgctacct cgacacgatg tacatcaccg 10560 cgcccggctg gggcgactcgggctccggcc acgtcggcga ggccggcgcg aacttcgacg 10620 tcaaggactg gatgtcgctctaccagggcg accagcagct cgactggggc aacgccgagt 10680 atctgcaggt tcccgggctcggtgcggagc gcctgcccta ccggctggtc ctcgacaaca 10740 accgtggcgc ctgggccaacccgtactcga cgcacacgct gaccgagtgg aacttcacct 10800 ctgcggtcac cagcagccagtcggcggtgt cactgccgct gatccagctc gactacggcg 10860 tggacaccga caagtccggccgggccgacc ggcatgccgg gctgacggtg accgcatcgc 10920 acctgcccgg caccactgccgcgatcgcga agccgtccgt cgaggtctcg tacgacgacg 10980 gtgcgacctg gcagcggaccggcctgagcc ggtgcggtga cggatggcgg acgaacctgc 11040 gagccccgcg gtcggccggcttcgccaccc tccgggtcgg cgcccgggac aacgcgggca 11100 acaccgtctc gcagacgatcacccgggcct tcggcctgcg ctgacgtcac aacccgggag 11160 gtcacctggg acagcaccgtgtgggtggga ccgcagccgg cggccccacc cacacggcat 11220 tcccagtcgt cagaggatctatgaggacac cccgtcgcgg aacggcaagc gagtgctcgt 11280 ggatcgcctg tggccccgcggaatgagcaa agcgcgggcc cacctggacg aatggatccg 11340 cgacgtggcg ccgccggcggcaggtcggcc aagtgcttcg ctgctgccgg agacgtggcg 11400 gccgacgcgc gccaggcgcgtccccggcca gaccattcgt acagccgccg gattcaccac 11460 gcactccgca tgcacccgttcaggcgcagc cggcgagggc gtcgatcgct tccttcccgt 11520 ggtgagtccg gccggttgggtcgggtcact cgggtccgcc gagcgggagc gtcggcgttc 11580 cgccctctcg caggcggtggtgagcgactc catggggtgc ggggtctgca cgaagacccc 11640 gcgttccggg tcgaggccgacactctggcc gtactcaccg actcgtagac cggttggctc 11700 tctgcgcctc ttcgggatgtgtcgcgcatc acatccggcc cctgtcacgt ccaagccgcc 11760 tgctcccatc ttgtgggcgtcagtgagcga ggcgaggagg acggaacgat gagcgcacgg 11820 caggagacca gcacggcgaccgcgcaccac gtggtgatcc tgggggcggg gtacgcgggc 11880 atgtccgccg cgatccagctcgcggcccgg gtcaaggggc gcgaggacgt acaggtgacc 11940 gtggtgaatg cgcaggagcggttcaccgag cggatgcgtc tgcacctgac ggcgaccggc 12000 cagcggctcg ccgagctgagcatcccggaa ctgctggagg gcacggaggc acggttcgtg 12060 cgcggctggg tgacggcggtggacgcggac gccaagaccg tgcggatcga cgacgaccgg 12120 gtgctgccct acgacacgctggtatacggg ttgggcgggg tggccgacac gtgcacggtg 12180 ccgggtgtcg aggaccacgcgtacaccctg agcagtgcgc aggacgccga gttgctggcc 12240 gtccggcttg ggcgtctcggcagcggcacc gtggtggtcg cgggcagcgg gctgaccggc 12300 gtcgagtctg ctgcggagatcgccgagcgg cacccggagc tggacgtcgt gctgctgggc 12360 cgggccgagc ccggcgcgaccatgaacccc aaggccaagg cgtacgtgca ggccgcgctc 12420 gaccggctgg gcgtccgggtgcgtgccggg gccgaggtgg tgaaggtgct gtccgacacg 12480 gtcgaactgg cgggtggggagagcatcgcc gccgacgtgg tcctgtggac gagcggtacg 12540 cgcgcgtcgc cgctggcggctgccgccggg ctggccgtcg atgagcgcgg acgcatcgtc 12600 accgacgccg cgctgcggtcggtgtcccac cctgacgtgt acgcagtcgg tgacgcggcc 12660 gcgatccgcc agggctacggcgtaatgcac ggcacctgcc agggcggcat gccgaccggg 12720 gtgcacgccg cgctgtcgatcgagtgtgtg ctcaagggca agcagcccaa gccgttccgc 12780 ttcggttact accacacgccggtgagcctg gggcgaggcg acgcggtggt gcagttcacc 12840 caccccgacg acagcccgcgccgaatctgc ctgaccggac gcatggcggt caggtacaag 12900 gagatggtga ctgcctccccttggccgacc tacggccgca tgaagaagat gcccgcctcc 12960 ggcgcgttct ggccccgcgggggccgctcc gcccgggtcc gggaggcacg gtgaccgacc 13020 cgaccgccca ccccggccagcaggccttcc acgagtaccg caagctgctg ttctccgtgg 13080 cctaccgcct cctcggcaccgcggccgacg ccgaggacgt ggtccaggac gcctggatca 13140 agtggtccgc tgcggaccgttcgcaggtgg ccgaccccag ggcgtatctg acgcggatcg 13200 tctcgaacct ggcactggaacggctgcgct ccactcggca caagcgggag acctacatgg 13260 ggccgtggct gccggagcccatactcaccg gcggggacgc ctccgaggcc gtcacggagg 13320 ccgagtcggt gtcgatggccatgctggtgg tgctggagac gctcagcccc ctcgaacgcg 13380 cggtgttcgt gctgaaggaagtcttcgact tcagccacgc cgagatcgcg gaggcggtgg 13440 agcgttccga ggccgcagtacggcaggccg cgcaccgtgc tcgtgagcat gtgcgggccc 13500 ggcggccgcg tttcgccacggaccggtcgc ggcagtgcga ggtgaccgag cggttcttcg 13560 ccgcagcgac cggtggtgacatcaacaccc tgatggaact gttgtcgccg gacgtcacgc 13620 tgtggaccga cggcggcggcaaggtccgcc aggccatgcg tccggttgtg ggcgcggcca 13680 cggtggccgc ttggttcgcggccatcggca ccgttgccta ccagggcgtc gagcccgccg 13740 acatgaaagc cgaactcgtcgagatcaacg gcggcccggg catggtgttc agcggactgg 13800 gccgtgtgat cgccaccgtcaccttcgact tcgacgccaa cggccgcatc accgccatcc 13860 acaacgtcgc caaccccgacaaactccagg ccattgccga cgggacagct cgcgacgtcg 13920 ggacgcgctg agacctgcgcgggatcgccg cggcccgttg ggcgagatcg tcgcccaaca 13980 gcccgaacgc aatcccgagccacccaccca cgacccggcc ggcccaggcg cagtcaccgg 14040 cggtcgaagc gcggggcgacaacacgacgc ctcggaatga aacaggagga gccagaccaa 14100 gacacccatg ccatcactgttcccgcatca cagaacacgt cagccccaac agccccgtcc 14160 aaccgcaagc ggatatcggcccgaacgggg ctaatgcaga gccaagatcc ctctgcgtgc 14220 gtgcttctga tcccagggatctggcctgcg gtgatcgata ggttgtacgc atggttgcga 14280 acgagggtgt ggccgtcgtcggggcggacg tcgatgaccg aggctggtcc gacacgcggg 14340 cttgggtgga aaagcgtgtaggcggagggt gccgcatcga cgctgccgag cggctgcgtg 14400 gcggatggac ttcggagatgcggcgtctac gcactggcgg aacgggagag ccgcagtgga 14460 tggtgctgcg gtcgttcgtgaagccattct tcgttcagca tgcgaaggga ctgctcactc 14520 gcgaagcgga aatcctgcgtctgctcagcg agaccgatgt gccggtggcg gagttccacg 14580 gtgtggatgc gacggcggtgtactgcgacc acccgtcgtt gctgatgtcg ctgctgcccg 14640 gcagtgtgcg gttgggtgaggaggaagcgg ggctgcggtc ccagctgctg gcccgccaac 14700 tcgtcggcat tcaccaggtgcgggtcccgg tcgaggcccg gccgcgcgtt taccaggcct 14760 ggacgtcgcc cgaccgtgtgcggctgcccg aatccaccga gcggctggac ttgtggcagc 14820 gggctgtgga catcatccggcgcgatcccc ctgactaccg gccttgcttc ctgcaccggg 14880 atttccaccc cggcaacgtgctcttcaccg ggcagggcgc ccgttcggag atcaccggtg 14940 tcgttgactg ggtggaggcgtcctggggcc cggcggatct ggacgtggcg cactgttcca 15000 cggccctggc tctcctgcacggcgccgagg agggcatgcg cctggctgat ctctatctgg 15060 cttccggggg cacgttgagtgaggacccgg ccgctcacct gtactggcgc gtcctggatg 15120 cactcgcgtt cgcaccggatgccgagaagg tcgcggtgcc ctggcgcgaa ctgggacgca 15180 aagacctgac ttcggacctgctcaccagca ggctggagga ataccttgag gagctcttcc 15240 gccgcttcgg ctgatcgtctccacctggcg gggcctgggc agcagactga gccacggccc 15300 gggaacggcc gcgcgagcggtgaacagtgg cctggccggc tttgacggct cagctggagg 15360 actcgtcttt cttgcccaactgccggtgcg cgggcccttc ttgtccaact gcccgtgcgg 15420 gggcctttct tgtccaactgtcggcgtgcg ggcctttgcg ggcgaggcgg cgggtcatga 15480 gggtgaagag tgaccaggtcacgtgggctt cgctgcgctg cggggaggcg ttcgtagccc 15540 cttgcgttgc ggcgggcgcgcaaggtccag ccgatcgtgc gctcgacgcg ccggcgaggc 15600 tcccccggcg ccccgcaccgcggccgctcg gtggtgacac gtgggcgacg gcctgacccc 15660 tacccgtcgg ccggccttcgtcagtccatg cgctgctgcg gttcggactg cttggtgccg 15720 ctctcggcga gagccgtgcgcgcggcctgc agcagttctt cggagatcgg gtcgccccgg 15780 gtggcgcggg cgagggcgagggcgccgacc atggtgcaga gctgggccat gccgtcgtcg 15840 tcgccggtgg ccagccgagcggcgcggttg cgtatcccgt tgatgtagac gtggcgggcg 15900 cgggcggcct ggtcgggctcgcgacccagg tcagcggcga atccggagac ggggcagccg 15960 tccgcggcgt gatcgcggtgccagaccgag aggtactgct cgatcagcgt ccgccgggcg 16020 gcctcgtgct ccccggcgcgctcctcgaac gtcgccgccg aatgtgccgc gtgctcggcg 16080 aaggcgtggg cgatggcctcgtcgaccagg tcgtccttgg acgcgaactg cttgtggaac 16140 ccgccatggg tgagcccggcggccttcatc aggtcggcga cgctgaccgc ggtgcctttc 16200 tcgcggaaca tccgggaggcggtggctacg acgcgctgcc ggttctcctg tgcttgcgcc 16260 tgggacacgc ggcccatggaatcactctcc tcaattggat gtcacttgcc atctatccta 16320 gccccgttta gattgtttgcgacatctatt tcatgtcgat gagacccccc gcaggtcagg 16380 cagggtcttg cgcggcctcccaaggttcga ggccgacggc ctccgcccgg caatcgcaga 16440 accaccgaca gcactggagaccccctgtgg catcgccacc caccccctcc gagtcctcga 16500 cgcccacgag gtccacgccgcccgtgcctc ctgcccagcg acgacgtgtt tccgaggcgg 16560 tccaccaccg tcgctgggccatcctggccg tgctcgcgtt cagcctgctg gtggtgatgc 16620 tggacaactc gatactcaacgtggcgatga agaccatcgc ccagcccgcc cccaccggac 16680 tcggctccac ccagagccagctggagtggg cggtcaactc ctacacactg gtcttcgccg 16740 gactgctctt caccgcgggcctgctcggcg accggctcgg ccgcaagaag gtgctgctct 16800 tcggcatgct ggtgttcggcgcgggatccg cgctgtccgc gctgtcggac tcctcggggc 16860 agctcatcac cttccgtgccgtgatgggcc tcggcggcgc cttcatcctg cccgccaccc 16920 tcgcgatcat catgaacgtcttcgagcgcg aggagcagcc caaggcgatc ggcatctgga 16980 caggtgtggt cggtttcgccatcgccgccg gccccatcgc aggcggcgtc ctgctggagc 17040 acttctggtg gggctcggtcttcctggtca acgtaccgat cgtggtcgcc gcgatcgccg 17100 cgatggcggc gatcgtccccgactccaagg acccccggcc cagcaggctc gacgcggtgg 17160 gcgtgctgct gtcgatcgtcggtctggccg tactggtcta cggcatcatc aagggcggcc 17220 agctgggcga cttcacccggcccgaggcat gggcgacaat cctcggcggt ctggcggtcc 17280 tcgccggctt cgtggtgtacgaggcccaca gcgaccatcc ggccctcgac gtcacgtact 17340 tccgcaaccg gcggttctccgcgtccgttg ccgccatcgg cctggtcttc ttcgccctca 17400 tgggcgtcac attcttcagcgtcttctaca accagagcgt gcgcggctac agcgtcctgc 17460 agtccggtct gctcgtcctgccgctggccc tctcccagat gttcttcgcc ccgagagccc 17520 ggctggtcgt cgaccgctacggcgcccgag ccgtgtgcgc cggcggcctc gccctgaccg 17580 ccgtggcctt caccggcttcctgctgctcg accaggacac ccccatctgg gtgctggagg 17640 tgctgttctt cctgatgggtacggggatgg cccatgtgat gccgcccgcc accgtcatga 17700 tcatgtcgtc actgccgcgggagaaggccg gctccggatc cgcggtcaac aacaccttcc 17760 ggcaggtcgg tggggccatcggggtcgccg tgctggggtc gctgctgtcc accacctacc 17820 gcaacggcat cgacggccacctgggcacgc tgcccgccgc cgaccgccac gccgcaggtg 17880 aatccgttca ggcaaccctggccgtcgccg agaagctggg accggccggc agcgcgctgg 17940 tgcggcccgc gcacgacgccttcatccatg ccatgcacat caccgcggca gggtcggcag 18000 ccgtcgccct ggtcggcaccgtcgtcgtgc tggtcttcct gccaggcagg aaggtcaccg 18060 cggccaccgg cggtgacaagtgagcctgcg ttcggcacgg caaccggcca ccggcggtgg 18120 cccggcaaag acgccctcctcatggacgtc ttgaagacca cggacgggcc gacgccggag 18180 ccggcgggcc cgccagatgctgcggccggt ctcctcgcag gcggacgggc tcaccgagcg 18240 gacaaccgag acggcactcgaaggagtacg cccgcgcggg tgaggccagt accgacttca 18300 gcgttgccga cctcctccgcccggcaatcg cagaaccatg gacagcactg gagacccctg 18360 agaaatcgcc acccggatcagccgtgacat cgccggggcc aacgccttgc tgccagagcc 18420 tctgacgctc tgatcacacctcaactcaga tattccatta ctccgaagga gatcgactga 18480 tgactgccga caacctcgttctcatccccg gtgccggcgg cgtgggccgc acggtcttcg 18540 agcacttgcg tgctcaggatgtgccggtgc gtttcatggt ccgtcgtgac gacgagcgtg 18600 cggctgagct tcgggcgctcggcgcggagg tcgtcatcgg tgacctgacc cggcccgaga 18660 cggtggctgc cgcactgcagggtgtggcgc ggatgcactt cgggataagt gtgtcgccgg 18720 accaactgct ggcggcgaccgtggtggcct ccgtggcgcg ggagtacggg gagctggagg 18780 ccctggtcga cctgtcgcagatgacggtgt cccagatgac cgccaccagc accgaggagt 18840 cgcaccagca gcggctgcactggctgacgg agcagatgct cgactggtcc ggcctgcccg 18900 tggtgcacgt ccgaccgacggcgttcctcg acaacccgct gttcaccacg atcgcggcgc 18960 ggtcgatcca ggcgaacggcaccatcgcgt tgccgttcgg caccggacgc acctcgccgg 19020 tcgccgcgga cgacgtcgcccggaccatcg ccaccgtgct ccgcgacccg gctccgcaca 19080 tcgggcaggt ctacgaactgaccgggccgc gctcggtcga catgacggag ctggcggcgg 19140 agttctcacg ggcgctgaatcgtccggtgt cttatgtcga cctgccgctg gaccggtggg 19200 aggcccagct cccgaagctggggatgtcgc cgcacgtcgc acagcacgtc gccaccatgg 19260 cccggctcca ccgggacaaccgttatgacc gcacggccga cggcgtcgag cgcgtgacgg 19320 gcacccccgc catgtcgatcgaggcgttcg tggccgcccg cagggacctc tacctgggct 19380 gagccgcccc gccagggccgtcccccgggg caactgccga gggcgctgag cctctcccgg 19440 cctggggtca gggacaggggccgagacggt gcttctgctg gtcggccgac ggcagaggtc 19500 cttctccccc cgcccgaagagcgggccggc cccacggagg gtgaagcccc acgaccgagc 19560 cgacccgcgg gagcagcactgcgacaggtg ttgtggtcag actgcctgaa ctgcgttcgt 19620 cggccggggt tgtgaggctgtcgggtggag aggcgcgcgg gtgaggcgtc gggtcatgag 19680 ggcgacggcg gcgggggtgagcatcgcctc ggcatgctgg ggcaggggct cgtcatcgcg 19740 ggtgccgcgc gtacgggtggtgcccccgcc cccgccgacg ggttcagtcg aggctgttgg 19800 tgaggatccg ttcggcgatgccgtcgcgca gggcggtggc gggtgagctg ccgggcttgc 19860 gttcggcctg atcgcggaagcaggcgagga actcctccgc gatgccggcg cgcgggtggc 19920 gggccaggac ctcggctttgaaagcggcgg agaagtcgtc catgtgacgt ccggagatgt 19980 cgaggctcgt ggagcgttccaggagatggc cctcggggtc cttcgtcacg tcgacctcgt 20040 cgagcatgtg gctgatgacgacctccgcga ggcgctcgcg tcgggactcg gaccagcccg 20100 cgccggcggc gaagacgcgggccacgtggg cgctggcgtc gtcgaagggg accgtgtggc 20160 tgtcgaattc cgggaccagggcgatgtcgt ggaacatggc cccgacgtag aggagttcgg 20220 ggtcgaaggc gatgtcctgctcggccccgt ggacggccgc ccagatgtat gaccggacgc 20280 agtggttcag gagggcgggcgagcagtagg cgcgggccac gtcggtggcg gccgtgcaga 20340 cgggggtgtc ggggagcagcaggtcgtcca gtgtcatggc gggcagtatt gccgaacggt 20400 ggcgtggggg tgccggggctcgagccgggt tccaccggat tcccagaatg gccgtgcagc 20460 gtgcgggagt tggggcgtcgggctccggtc aggcagtcgg ggacggcccg cgcgggcggc 20520 tcacgagtgg aattcgagcggtgcgtgaaa cgctccgggt gccccgtcgc gggctcagcg 20580 gcacggccgc ccgtggacccccggtccagc aggtgggacg gcccgcagtc cgtgcacccg 20640 gatccgtctc aggaggacctcttgaccacg ctcacgttcg ccgaggcgtc cgcgcactcg 20700 ggcggggatc tgcggtacctcccggccggg cacggggagg tgctggatct gagtacgtgt 20760 gtgacccgct ccggcccctcgccgcgcgcg ctggcggccg ccgccgcctt cgacccgtcg 20820 tccctgacgg tccatccgtacggtgccgac gaccgcttcc gtacgacgta tgccgcgtat 20880 ctgggcgtgg acgcacggcagttggccgtg acgcggggaa tcagtgagcc cctgtcggta 20940 cttgcgtcgc tccttccggccgagcggtcg gcggccctga cccccgacta cacgggcacc 21000 attcggcggt gggcccgtcatctgccgccg gccccggacg agcgcgactg cgtggactcg 21060 cgggtgcggc gtctggacgcggcgatgcgc cactacgcct acgtggtgtt ctcgaacccc 21120 aacaatccgc tgggcctgaccgtgccgcgc gacgttctcg cggagctgct gtcccggcat 21180 ccgcgctgca cgctggtggtcgacgaggcg tatgtcgact acatgggcga ccggcggcag 21240 tcggcgatga cactgccccaccgcaatctg gccgtgctcg cctcgcccgg caagccgttg 21300 gggatcgccg gtacgcgcacgggagtgctg tggacgcccg atgacgtgct gcgcgagcgc 21360 gtggcgcgac ggctgcccgagtggccgctg tcggcgtacg acgcgcacgt ggcgtgtgcg 21420 gcgttgcagg acaccgcctggatcgcctcg gcgctggagg gggcgcggga ggacgcccgc 21480 cgtctggagg ccgttctggtccgctgcttc ggggacgcgg tggtcaccgg cgttccggtg 21540 cactaccggt tcgtctacgacgagcgcccc gagcggcttc acgcgcacct gctggccgac 21600 gggatcgcga cacgactgctcgtcgacgac gagccggggc gggtgcgggg gctgcgggtg 21660 ctggcgccgg gcagcggggagctggcccgt ctggaggctt cactgaacgc gttcgtgcgg 21720 tgactggcgc gaggttcaacaggcttgcgc agcagggaca tcggggggat tcgccgggcc 21780 cgtcgggcga tcggcgcgccgggtggagcg cggccgctgc actcccggca cagggcccct 21840 tgagagcggc ctgaggactccggtgtgggg ggcggggcgc ggctagggtc ccgggtcagt 21900 caggtgcacg tccctcgacttcttctcgtg ttcggcgtcc tcgtcccggc gggcgccctt 21960 ctcctcggtg cacagcccctggaagcggtc ctcatggaat tcagctcgcg atcgtcgacc 22020 tcgtccacga ccggtgccccgctgctctcg gtgcgcagcg ccggggagat cgtccccgtg 22080 ctctgtccgg acggcttcgaagtgggcggc gggcctcggg tcgtggaggt cgtcgccgac 22140 cggtgggcgg gcaagtcgaccgtgctggac gggctgtccg cggccctggc gggccggggc 22200 tggacggtcg cggccggtgcggcgtcgccg gcccggtccc gtgcgccgtt cggggtgttc 22260 gtggaggctc tcgaggagctggtggagcac ctggatcccg atcggctcgg cgggcgcggc 22320 cgggatcacg cggcacagctcgccgcgctg ttcccggcgc tctcccctga cgcgccgcgg 22380 gacacgtccg tggacacgtaccggatctgc cgtgccatgc gtgccctgct ggagatgctc 22440 gccgccgacg gaccgctgttgctggccctg gacgacgtgc actggatgga cgccgattcg 22500 ctggaactgc tcgggcagctgctggagcag ccggtcgcgg ggccggtcgt gatcgcggtc 22560 gcccatcggg ggcgtccgggcgatcgtgtg ctgaacgggc tccttgcgcg gtgcgcctcc 22620 cggggcgggt gtctgcgggtgcggcccgcg ccgctgacgg agcaggaggc catcgcgctg 22680 ctgccggacg atctgagccgggtgcactgc gagacgctgc tggtggagtc cggcggcagc 22740 ccggggctgc tgcgggccttcgccacgctg cgtcaggtgc cggggctcac gggtgcgtgg 22800 actccgccgt tgccggcggatctgctcgcc gatgtgctgc gtgacttccg tgcgctgagc 22860 gccgagggct ggcaggtggtgcgggccgcc gcggtgctgc acgagccgta cgcggccgac 22920 gatctcaagg agatcgcgcacgtcgacgac gcggcgctgt ggcgtgctct cgacgagctg 22980 atccatgagg aactgctgcggccggacgag ctgccctggc agctgcggtt cgccaatccg 23040 ctgctgcggg tcgccgcctaccagtcggcg ggccggggct ggctgctcgg cgcgcacgcg 23100 caggccgcgc ggatcctcgcgcggcccggc tacccctcgg cgcacatggc caggcatctg 23160 gagcgtggcg cggcgctgcgggacgagagc ggtgcgcggg tgctgtggga cgcggcggca 23220 cggctgctgt ggcaggacccgtcccagtcc gtctcctgga tcagggcggc gctggcgtcg 23280 cgggcgggcg gtgacctggagccgcgcgat cggctgctgt tcggcaaggc gctggctctg 23340 gcgggccggc tcaccgagtcgctcgccacg ctggggccgt tcgggcaggg cggggtcgag 23400 gcagggctgc gggtggagcgggtgcggtgg cgggcgcatg cgcaccggct cctcgggcac 23460 accgaccgcg cgcaggacgaactggagtcc ctgctggccg agttggagcc gggggaagag 23520 gggctgcggg acgtggtgcggcacgcgttg atcgagctgg cgctgcacgg ccgccgcccg 23580 gtgccgccgc cgcccggtgggagcgagccg gggatgcccc agggggcgac gggggcgctg 23640 cggggcaggc tgctggcgcagctggccgtc gcggcctgcc ggggcgggga gcaggagcgg 23700 gccgaacaac gagcactgacggcacgccag ttgctggatg cggcaccgga cggcgaggtc 23760 gccgagcagc tcgagggcctgtactggctg gcgaccgccg aggcggcgct ggggcggccc 23820 tcagccgcgt gcggccactacgagcgcgga ctgcggatcg ccgagacccg ccggctcatc 23880 gaccacgtcc cgctgttcgcgatggcgctc agcgcgctcc agctgcacac cggtgacggg 23940 gcgggagccg cccggcacgccgcgtgtgcc gagaccgtcg ccgcggcgac ggagagcgag 24000 cacctgcggg agcgggcgctgcggctgcgc agggagacgg cttcgggggc cggcgtcgcc 24060 gtactgcccg gccagcggcagccggcggac gcggcggcgg tggccctgga ggggctgagc 24120 cggcgcgaga gtgaggtggcgctcctggtg agcgggggcc gtaccaacca gcagatagcc 24180 cgtgtcctgg agctgagccacaagaccgtg gagacgtatc tgggccggat attccagaag 24240 ctgcaggttc agtcgcgggccgaggtggcc gccatggtgg ggcgttcgga ccggctcggc 24300 gtggccgcgg gctcctgagcggggccgttc ggcgcgggcg gtcgctcgtg gtcggccggc 24360 ggacggcggg tcgggagatctcctgacacc acaggaaggg ccggatggtc caggttggga 24420 cggtccggac acgggaggggcggtcgtgga cgtggcctct ccatcaccac ggatccatca 24480 ccacggacac atcaccacggttccatcacc acggaggaag cagatgagcg atcgggacag 24540 tgccgacatc gcgtacgcggtggtcgtcaa cgacgagcag cagtactcga tctggcgcgc 24600 cgacgccgag ttgccgcccggatggagtgc cgaaggcacc accggcacgc gcgaggtgtg 24660 cctggcccgg atcgacgaggtgtggacgga catgcggccg ctgagcctgc ggcgcgcgat 24720 ggcgcaggcg ggccgcagctgaggccgtcg gcgcctcatc gacatctgga ggactcaatg 24780 cagacccagc tctcccagggcgctcaactc tccgccgagc agcggaagtt gctcgatcag 24840 ctgctggacg cggacggtatcgaccgggtc gccggcaagg tgacgcggcg caccggtgac 24900 ctgtcccgcg cgccgctgtcgttcgggcag gagcggctgt acttcgtgga caagctgcgt 24960 ccgggttcac cgatgtacgtcggttccggg gcgctcaggc tgcgcggtgc gctcgacgtg 25020 gaggtgctgc ggcgctcgatcgcgctgctc gtggaccggc acgaggtgct gcgcaccggt 25080 gtggtggagg acggcgacggtgtcgagcag cgcatccggc cggccgggac ggtccaggta 25140 gagctgccgg tccggggtgccgcgtccggc gaggtggccg accgggtgcg cgaggcggcc 25200 gcggagggct tcgacctgtccgcgccccca ctgctgcggg ccgtgctcct ggagatcacc 25260 gacgcggcgg agccggagtgggtgctggtc ctgtccgtgc atcacatcgc cgtcgacggc 25320 tggtcgctcg gcctgctgatgggcgagatg agcgagctgt actcggcatt ggtacagggc 25380 gccgagcctt ctctcgccgaactccccctg cagtacgccg atttcgcctc ctggcagcgc 25440 ggctggctgg agcagggcatcatcgccgat caactcgcct actggagtga ccagttgagc 25500 ggagtagagg cgaccgaggtgcccagtgac cggccgcggc ccgcggagcg cagctacctg 25560 ggtgacaccg tgccgctcgcgctggacagg ccgctgacgc agtcgttgcg ggcgctgacc 25620 gacagcggac agtcgaccctgttcatggcg ctgaccgcgg tgtgggcaag cgtgctcggc 25680 cggttcgcgg accggcgcgacgtggtggtg ggcactccgg tggccgggcg cacgcgcgcc 25740 gagctggaga gcgtcgtcgggttcttcgtg aacacgctgc cgctgcgggt ccgcatcgag 25800 gacggcgaca ccttccgtacgctgctgctc cgcacccgtg aggtgtgcac ccaggcgtac 25860 gcccatcagg acgtgccgttcgagcggatc gtggcggagc tgcagtcggg gcgcgacgcc 25920 tccgggcaga cggcgatcgcccggcactgg ctggcgctgc acaacacgcc cgcccccgat 25980 ctgacgatgc cgggcctcatgagcgaagcg ctgcccgcgc tcatcggcac ggtgcgctgc 26040 gacctgtcgg tgcaactggcgccggacggg cagggcgggc tgagcggccg cctcgagtac 26100 tccacggaac tgttcgaccgggccacggcg aagaccctgg catcggcttt cgagacgctc 26160 atcgcctcgg tggcggccgcaccggacagc ccgctggacg agctggacat gctggccccg 26220 caggcgcgcg aacacatcgtgcgcgacctg tcgggcgctg cccgaggtgc cctcaccggc 26280 ccctgggtgc cggagcggttcgccgcgcag gtcgacgccc accccgacac ggtggcggtg 26340 gtcgaggccg acggcgggcagctgacgtac ggccaactcg acgtgctcgc caaccgggtc 26400 gcgcacctgc tgacggcgcgcggtgtcggc gccgaggacc gggtgggtgt actgctcccg 26460 ggcggcggcg cgctcgcggccgccgtgctc ggcgtgctga agtcggcggc ggtggtgctg 26520 ccgctcgacc ccggctacccgcccgcccgt ctcgcccagc tggtcgagga cggcgcgccc 26580 cgactggtgc tttccggtgccgggtcgacg gatctgctgc ccgacctgga cgtcgtggac 26640 gtcacgggcg aagaggtcgcgggccacccc ggacaccgcc cgccggccgt cgcgacggcc 26700 ccggacgccg gtgcgcatctgctgttcacg tccggcacgt cgggccgccc caagggcgtg 26760 ctcaccaccc gtgccggcctggtcaatcgc ctcatgggca tggagcggca gttctccgtg 26820 ggtgcgggtg accgggtgctgtgcaaggcg ccgctgggct tcgacgtcgc ggtgtgggag 26880 ctgctgctgc cgctcgtcaccggcgccacc gtggtgtcgt gccgggtggg cgggcaccgc 26940 gacctggagt atctgcgcgagctgatcgac cgccacgggg tgagtgtctg ccacttcgtg 27000 ccgtccttgc tcgaggagttcgcgaacgcc ccggccggtg cgcacccctc gctgcgtctg 27060 ctgctcagcg gcggcgaggagctgccggcc ccgctcgcgg aacggatcct cgatcggttc 27120 gggcacgccg agttcgtcaaccagtacgga ccgaccgaga ccgtcatcga cgtcacctcc 27180 gggcgggtct cggggcccgtcccgcaccgg gtgccgatcg gccggccggt gcccggtgcc 27240 gaggtgtacg tcctcgacgcgtcgatgcgg ccccaacctg tgggcgtcgt tggtgagttg 27300 tacgtcggcg gggtgcaggtggcccgcggc tacgtgggcc gtgccgcgct gaccgccgag 27360 cggttcgtgc cgcatccgttcgcgcacggg cagcgcctgt acgcaaccgg ggaccgggcg 27420 cgcctccttg cgggcggctcggtggagttc ctcggccgga ccgacgacca ggtcaagatc 27480 cggggcaacc gggtcgagcccgcggaggtc gcggcggtgc tgcgcgccca cccgcaggtg 27540 tcgggcagcc acgtcaccgtgcagcgcacc ggcgcggcac cgcgtctgac cgggtacgtg 27600 acgtcggacg ccgcgaacgcggacgaactg acggaggagc tgcgtgagtt cctgcgcgag 27660 cgggtgcccg aggcgatggtcccggcgcag ctcttcgtgc tgaggcagtg gccgaccggt 27720 gcgcacggca agatcgacgtgacggcgctt ccgcagccct cgggcgaacg tccgggcgca 27780 cagcgagagt tcgcggcgccggtcactgac gtgcaggtcc gcctcgccgc gctgtgcacg 27840 gagctgctcg aggtggagcggatcggtctg acggactcgt tcttcgacct gggcgggcac 27900 tcgctgctcg cgatccgggcgatctcccgc atccgcaagg agttcggggt ggggctgcag 27960 atcggccagt tcttcaaggcgccgacgctc gcgggcctcg ccgaggcgat cgaggcgaag 28020 ctcgcggccg cggacgggaagccggccgga ccggccccga tccccaggat cgaccgcggc 28080 gcgcagcgcc gctgaccggccgccacccat cgtgagggag actgagcatg accattgagg 28140 tgcacgacgc cgtggccgtgagcccggtgt gggagggccg ttccctgccc gcgctggtgc 28200 gctgcgagag ccccgggggggcgctgcccg cctgggtgag cgcccatcgg gacgaggtcg 28260 acgcactggc acgcaaggccggggcggtgc tgctgcgcgg cttcgacgtg cgcggcgcgc 28320 aggacttccg ggccgtgatggacgccctgt cgccgcaggt gctcgcctac ggcgagcggt 28380 cgtcgccgcg cagccaggtgtccgacggcg tctatacgtc caccgagtac ccggcggacc 28440 agccgatcct gctccacaacgagcagtcgt acacggccaa gtggccgaca cgcatcgtct 28500 tcttctgcga gcgggcggccaccgagggcg gccgcacacc gctcgcggac tcccgccgga 28560 tcctggcgcg cctgcgcccggagacggtgg acaggttcga gcgcctggga gtgcgctacg 28620 tccgcaacta cctgccaggcatcagcctgc ggtggcagga ggcgttccag acggaccggg 28680 tggaggacgt cgccgcctactgtgcgcgtg ccgacatcac gcccgagtgg gtggacgagg 28740 accatctgcg gaccgtgcaggtcaggcccg cggtgcgcag gcatccggtc accggggagc 28800 gcagctggtt caaccacgcgctgttcttcc acgtgacgtc gctcgacccg gaggtcagcg 28860 ccggtcttct ggaggccctcgacgaggagg acctgcccta caacacgtac tacggggacg 28920 gttccccgat cgagagcgagacgctcgcgg agctgcgggc cgcctacgcg gccgagacca 28980 cgggcttcga ctggcagcccggcgacgtgc tcgtcgtgga gaacatgctg gtcgcacacg 29040 cccgtgagcc cttcgtcggcccgcggcgca tcctgaccgc catggccgac gccatcgacg 29100 ccggcgaggt gcccgcatgacgctcaccga cctgaccccc gcgctggagg acggcacggt 29160 cacggtccgg ccgctgcacgaggagccgtt cctgctggag atcgaggcgc gtgagccggg 29220 cctcgacctg ccgcggtgggtggccggccg caaggaggag ctgcgggcgg gcctcgaccg 29280 ctgcggcgcg ctgctgctgcgcggcttcga cgtaccggac ccgcaggagt tcgggcgggc 29340 cgcccgcgcg ttctcgccggacctgctcgg ttatctggag cgtgccgcga cccgcacgga 29400 ggtcgccgac aaggtgttcacctcgacgga gctgggcgag gaccagtgga ttccgttcca 29460 ccacgagatg tcgtactcccacaactggcc cggcctgctg tacttcttcg gcgatctgcc 29520 gtcgccggag ggcggcgccacaccggtggc cagcgagcgc cgggtgttcc ccctgatccc 29580 ggccgaggtc cgggagcgcttcacacggca cggagtgcgg tacgtgcgca actacggccc 29640 ggacctcaac gagtcctggcaggtggcgtt ccagaccacc gaccgggccg aggtggaggc 29700 gtactgccgg cagtcggacaccgcgttcga gtggaccggt gacgacgagc tgcgcaccac 29760 cgcggtgcgc caggcggtggcgaggcatcc gcggaccggc gagacggtct ggttcaacca 29820 cgcccatctg ttccacctgtccaacgtgga gccggaggtc gccgagatcc tcgtcgagga 29880 gtacggcgag gagggcctgccgcgcaacgc gtactacggg gacggcgcgc cgatcgagaa 29940 cgaggtcgcc gagctgatccgatccctgta ccgggaggcc gccgtgtcgt tcacatggcg 30000 gcgcggcgac gtgctcgtggtggacaacca cctcgccacg cacgcccggg agccgttccg 30060 cggggaccgc cgcatcctcgtcgccatgtc cgacctgcac accgagctgc cggccggccg 30120 atgacggctg ccgcgggccgtcaccgcacc attcgcacca ccggaggaac catgctgctg 30180 cgccccacca ccgacgccgacctcgcccgc gtcaccgact tcattgtcga ggagccgatc 30240 agctggattc cggccgaccgctaccgggcc gaactcgccg aggggatgta ccggccgcag 30300 tggtcgtgga tcgccgaggcggacggcgac atcgtggcgc gggccctgtg gtggggccag 30360 gccgcgagcg agcacccggtggcgctggac tgtctgcacg tggcgccctc ggtgcccgac 30420 agggcgggcc tcgcggccgatctgctgcgg gccggacaca cggcgttcgg ccaggcgggc 30480 gccgcgcagc cgccgctgtacaacctgacg ctgtccaacg gctggcgtga cgaccccgcg 30540 gtgtcggccg cgctggcctggcggcgcgaa gcggccctcg cggccggtct cacccacgag 30600 gtggaacggc tgcgcctggagtggacgccc gggaccgggg tgccggccga gtcggggcgg 30660 ctcaccttcg aggaggcgcccgacgaggag ttcctcgacg ccttccgccg ggtcgccgag 30720 ggcagcctgg acgccgagacccgtcagcac ctcgcggcgc acgggccctc ggccacggcc 30780 cgcaaggaac tggacttctacctcgggtgc cccggcgagc gcgcctggtg gcgcctcgcg 30840 cgcacgcccg acggggccgtcgccgggttc gcgatcccct cggccacccc gtacaacccg 30900 aacgtcggct atctgggcgtggttcccgaa ctgcgcggcc ggggctacgt cgacgtgctg 30960 ctcgccgaga tcacccgcagccacgcggag cgcggcgccg aactgatcac cgccaccacg 31020 gacacgggca acgcgcccatggcggcggcc ttcgaacgcg ccggttaccg caacacccag 31080 atccggatgc tcttctcggccccggccggc tgatccgccg agtcctcctg gagtacgtca 31140 tgacgcagac cgcccccttgtccccgagag accgcccggc ggcgaccccc gagcccgcga 31200 gccgccccta cgcgcgcggctaccagtccc ccgccgcact gcgccccgtc ctgcgcgcgg 31260 cccatcacac gcggctcgccaccacggcgg cggtcgccgt cgcggaccaa ctcgtgctga 31320 tcgccgcgat actggcggcatcctggccgg gctggtggtc ggcgcccgcg ccggccgcgg 31380 ccggcgtact catggctctgaccgcactga tcctcacggg acgccagata cgggcgctgg 31440 agtgcctggt ccacgaggccagccacttca actggagccg ccaccaccgt cgtgccaacg 31500 acaggtgcgc aacggtcctggcgggcctgc ccaccggtat ccgcatcgcc cgctaccggg 31560 ccgaacacct ggtgcaccacagcaggctcg gcaccgagga cgaccccgac ctgcagcgct 31620 acgcggagct ggacatcaccggcctggacc gcacgagccg ggtgggctac gcccgttccc 31680 tgctggtgcg gatcgccgcccaccaacgcg gctggtaccg gtccacgtcc acggacctgc 31740 cggcgctgct gctgccggtgggctgggcac tgctggtcgt gtcgctcccc gcggctctcc 31800 tgtggggcgc cgccgcggccgcggtggccg ccggggtgtg gctgaccggg ttcgcgctcg 31860 tcctgcccgt gctgcggttcatcggggagt ccagtgagca cagctaccac gacgcggaca 31920 ccgtcttcga ctccaccatcaccaacatcg gcccgctcca gcgccggctc atccatccgc 31980 acagcgacgg ctaccacaccgtccaccacc tgtggcccgg tgccccgcat caccaactgc 32040 ggctgctgca cgacgaattgatgtccgccg acccgcaggg attcgccggc cggctgcgca 32100 cccgcagccg catcgtggccgaccccaccc cccgtaccgg ccccggcagc acgccgtgga 32160 cgtgaccccc tgacacctgcctgaaccacc ttcgaggaat gagacccgac atgagtgagg 32220 tcgcagtgga gcagttgaagtttcttgcaa cggacgacgt gcgcaccgtc gccgaggagt 32280 tcggcacgcc ggtgttcgtgtacgacgagg ccaccgtgcg cgcctcgtcg aagacgatgt 32340 ccacccttcc caacgcgttcggcctgacgg tgcgttactc cctcaaggcg tgccccggcc 32400 aggcgatcat ccggctcttcgaccgggagg ggttctcctt cgacgcgagc tcggtgtggg 32460 aggcccgccg cgcggtgctcgccggggtcg agccggggcg catctcgatg accgcgcagg 32520 aggcggtctt cgacgagaacctgcgcgacc tcatcgacca ggggctgcag ttcgacgcgg 32580 gctcactcca ccagctcgaggagtacggcc gcgcctaccc gggcggtgcc gtcgcgatcc 32640 gcatcaaccc gggcttcggctcgggcctgg tcggccgcct caacagcggt ggcccgcagt 32700 cgagtttcgg catctggcacgagcagctgc ccgaggtcaa gcagatcgtc gagcagtacg 32760 ggctgcgcct ggtgcggctccagtcgcaca tcggctccgg ccaccactgg gacatcctcg 32820 tcgacgcggt acgcaccatgctgtcgttcg cccgcgactt ccaggacgtg cacaccctgg 32880 acctcggcgg cggctaccgggtctcggccc tgcagagcga cccggtctac gaccacaccg 32940 agtgggcccg cgtcgtcgccgacgagctca agcagttcgc cgaggagacc ggccgccggc 33000 tgcacaccga ggtggagcccggcacctatc tgacggcgct ggccggctcg ctggtcacca 33060 aggtcatcga caaggcggacaccggcgcgg aaggcaactc cttcctcaag atcaacagtg 33120 gtctgaccga gatcgcgcgcccttcctact acggcgtcca gcacccgatc gtgtcggtcc 33180 ccgccacggg cggcgccgagcgcgcggccg acgaccgcga ggagttcaac gtggtcggcc 33240 actgctgcat cgccggtgacgtgctcaccg cgaagcagga ggagctggcg ccggtgccgc 33300 tcggccgtac ggagatcggcgactacgtcg tcatcgagcg ggcgggcagc tactgctcct 33360 cgatgtcgat gaagaacttcaactcctacc ccgaggccgc cgaggtgctg cgccgcactg 33420 acggcagctt cgacctgatccggtcccgcc agagcatcga gcaggtcatc gtcaacgagc 33480 ggatccccga gggtctgtgacgtctcgcca gaaaagcaca gacgctcggc cgggggtttc 33540 cccggccgag cgtcttgcgtggggctgctg agatccgagt cttcaggccg cgccgcccgt 33600 cagagcacgt atcggcgggagaccagcgtg agtcccgagt cgtccgcgga ccaggtcacg 33660 tcggccgggc gcgggacgctcagcgtgccc ccgggcagcc ccggggtccg ggtgacggcg 33720 cgcagcgtca gcggctgcgttccgcggagt gtcagttcca cccgggtgcc cggcccggtg 33780 ggcggggcgg cgaggagcaggccccagcgc cagccgtcgc tcgcgaaggg ccggttggtc 33840 ccgccgggga cggactgtccggccacctcg gcgcgcacca cggctgcgcc gccggcgtcc 33900 gcgtacaggc cgatctccgcggccctcgcg tccgtggcgc gcacggacag gatgatgcgg 33960 cggacgtcgc ccgcgcgggtggtggagacg accttcacct cggggacggc gacggtcgcg 34020 gcgggcgcgg gcccggtgtgccagcgggac ggctggtcga gggagggaaa ggtgctctcc 34080 agggacgtcg tggcgccgccgacgtagtgg tcgagccagg cggagtctcc cgctccgctg 34140 gaggcccagc gggcccggccggtgtcggcg tccagcgcgt agaggaggct gacctgcgcg 34200 gggcgggacg cggtgggcgcaccggcgagc agggccgcgc cgagcgcggc gagagcggct 34260 gccgtgccga ggccggtgcctgtgacgagc gagcggcggc gggtcttggc gggcaccgcg 34320 ggcagtgcgg tggcgagcgcgagggcgagc aggagcgccg gggcggcggc gaaggcgatc 34380 ggcaccgtgt cgaagaggagcgcgcccacc ggcaccagca gggcggcgca cggcacgagg 34440 ggcgccgtgg ccagtgcgccgcgccagggc gagccggggg cgatgcgggc cgcggccgcg 34500 agtcccgccg ccccggtgagcgcgggccac aggaacaggt acgaggcgcc cggcagccag 34560 acggcgctca gcagggtgcacagggtcagc cacccggtga ccgcggcgcc cgcctccaga 34620 gcgctgaccc ggcggcgcatgacaaccgcc cacacccaca gggccgttgc ggtcaggagt 34680 gcggcggtca ggtgcagcggtcctgccgtg taggggctgc ccagcgcgaa cccggtccac 34740 tgcggcctga gccaggtcgcgacgggccac accgcgctga ccgccgccgc cgcgcccagc 34800 agggcgagcg gcagtgaggccgcgctcagc gccgcctttc gggcccggac ggctccgcgc 34860 cggcgcgcgt accacccggcggcgccgagg gcggccacgg cgaggagcgc gccgggcagc 34920 accaggccca tcgggtagcggacgaggacc ggccccaggg tgaagtacgt ggccccgccc 34980 gcgtgcgacg tgccgctcaggtcggcgccg cccagccggc gggccgcggc caggacggtg 35040 tcgcccatgt cctgcaggctcgcccggctg aagtgggcga ggtcgtcctg gggggtgtgg 35100 tagttcgcgg aggtgccgatcacggcgaag ttcatgccgg tcatgccggc ctcgcgcagc 35160 acggtgaagt cggtgtcgttgggcagcatc cggtacacct cgtcggacag cgaggtggcc 35220 accgggaccc ggtcgccgagcgcggggacc acggcggcgt tcccggttcc cgtctcgaac 35280 atgaccgccc gtcccgaggtgccccgcgcc tccaggttga ggaccacggt ggaggccgcc 35340 ggtgccgtgt cgcgggcgacgaaggcgcgg gcccccagca ggccgggctc ctcgccgtcg 35400 gtgatcagga aggtgacgtcgttgcggggg gccggacccg tcttcagggc gcgggccacc 35460 tccagcgcgg tggcgaggccgatgccgtcg tcggaggcgc cgtggctgtt ctccgcagag 35520 tcgtagtggg cgacgatcagcaccctgccg ctggggtggg tgccgtgcac ggtggcgctg 35580 atgttctgtg tccaggcggtcacggccgcg ccgttgccgg tgtcgctgct ggtgcccggc 35640 tcgacgcgta cgccgagccccaactcgcgc aggcggcgga tcacttcgtc gcgtgcgcgg 35700 tcgtgggccg cggtgccggtggcgtgcggg ccgcccgaca cggccgccag gtgcgggtag 35760 gcgcgcgcgg cggagaaggagcccgccggt gccgacgccg gcgccggggc gggggtcagg 35820 ctcgtccaga gcacgccgacgagagcgacg gcgagggcga gcaccggcac cagaaggcgg 35880 gtgagggccc gcatcgccggtgacgggcgg ggcgggcccg gtgcgggtgg ccgcggttcc 35940 tccgcagcgg tgaggtccgcgggggtggta cgggtgtcgg ccatcggtca gctgcttcct 36000 tgtcgtcggt gtgctcaggtccgttccggt gcctcggtcc gttcggccgc cacggcctgt 36060 ccggtccgcg gggcgggcctgtcgtccccc gccgcgtcga gggctgccgg ggacagccgc 36120 agcagcgccg tcacggtggccgcggcgagc agggcggccg cgaacaggaa cggggcgcgc 36180 agccccgcgg cttcggcgagcgcgccgccg gccacgccgc cgagcggaga cactccgttg 36240 acgacgaacc ggtagccgccgttgacccgg ccgaacagtt cgccgggcac cagcgactgc 36300 cgcagcgagg tggtcaccacggtcgccagt gagatggcgg cgccgacgaa ggccagggcg 36360 ccggcggcga gcagcgggagcgaggtgagt ccggcgatca ggaacggcaa cgggcacagg 36420 gcgaaggcca gttggagcgtgcgggcccgg ccgatggccc gggtgacgag gggggcgagc 36480 agcacccccg ccaggccgcccacggcgagg acggccagga gcaggccgta ggcgcgctgg 36540 ctgatgtgca gcacgttcagcgcgtacagg acggcgatgc tcaggaccgc gaccacggcg 36600 aagttgagcg cgccgaccagtacgcacacg gtgcgcagca ctcggtggcg ccacagccag 36660 cggatgccgt cggcggtctcggccaccacg gcgcgcgcgg tcagcctgcc cgccgcggtc 36720 ggcgcggggc gggcgcccgacaggcggatg ctgaacacca ggaccgcgga gacggcgaag 36780 gacagggtgt ccacagcgaacggcaccggt cggccgagtg cgaacagggc ggtgcccagc 36840 ggcgctccca ggaactcgctgctcaccagc tgaccgccca tcaggcggct gttggcagag 36900 tccagggagc cgccgccggtcacgtcgggg acgacggcct gcgacaggtt ctcggcgagc 36960 gtcgtgatcg aggtgagcaggaacgagacg acggcgaggg cggtgatccc ggcccggtcg 37020 gtcaggatca gcaggaccagcgcaccggtg accaggaccc gggcgacatc gcagatccac 37080 agggtgcgcc accggtcgaggcggtccacc agggcgccgg cgaacggccc cacgcaggtc 37140 cacggcagct ttcccgccatcgccaccagc gcgaccgcga aggggtcgtg ggtgagggtg 37200 gccgcgtaga cggcgagggcggcggtgcgc gtgccgtcgc cgaatccgga ggcggcggtc 37260 gccgcccaca ggcgcccgtacccgtacgcg aatcgcggcg tgcgcggcgc cacgggtcag 37320 ccctccagct tctgctgcacctcgttgtgc gacatgctct gcacctgccg cacgagggcg 37380 gccacccggt ccacgtcgatgcctgcctcg gctccctcgg cgcgcatcag cgtggcgagg 37440 gtggcgacgg tgggcgcctcgacgaagaag tgcaggggga gttcgagccg gaacagttca 37500 cggatgctgg agacgacgatggtggcgagc accgagtggc cgccgcgttc gaagaagtcg 37560 tcgtcgaccg agacgtcggtgatctccagg gtctgctccc agacaccggc cagcacgtgc 37620 tgcaggtcgt ccgcgggagcccggcgctcc tttgccgggc cccggcccgg ccggggcagt 37680 gaagccctgt cgatcttgccgtacggggtg agcgggaaag cgtcgagcgc gacgaagaca 37740 tccgggacca tcggcgagggcagcgccgcg cgcagatgtg tgcgcagggc tggggtgtcc 37800 ggcgcgctct gcccgggcgccacggtcaca tagccgatca ggcgggcgcc gccgtccggg 37860 gccgcgtcgg cgaggacgagggcgtcggcg acctgggggt gggcgcgcag cgcggtctcc 37920 acctcggcgg gttcgacgcggaacccgcgg atcttgacct ggtggtcggt gcggccgagg 37980 aactccaggg tgccgtcgggacgccagcgg gcccggtcac cggtgcggta gacgcgggcg 38040 ccggggacga aggggtgcgggacgtagacc tcggcggtct gggcggggcg gccgaggtag 38100 ccgcgcgcca cctggacgccgcccgcgtgg agttcgccgg gaacgccgac gggctgcacg 38160 cggccggcgc cgtccaggacgtagaggtcg ccgcccggca cgggacgtcc gatgggcacg 38220 ggggatgcgg cgggcgcggtgacggtgtgc tcggcgatgt tgatggtggt ctcggtcggc 38280 ccgtaggagt tcaccaggcggatgtgcggt acctggagga ggagttggtc ggcgagatcg 38340 gccggcagct cctcgccggcgctgatcacc agcctgaggg cggcggcgcg gggcagcccg 38400 ccgaggccga tggcggggcgcagcgccgag ggtacgaaga agcaggtggt gacggcgtgc 38460 cgggtgagca cggagtacaggtagtcgacg tcggtgccca ggcccggggc ggcgagcacc 38520 acgcggccgc ccgaacggagcgcgaccagg agttcccaca tcgaggtgtc ggcgctgagc 38580 ggggtcttgt gcagcacgcgttcgtcggcg cccaggccga ggtccgcgca catgccggcc 38640 agccggctgg ccagtcccgcacgggtgatg agcacgccct tgggggcgcc ggtggagccg 38700 gaggtgtaga ggacgtacgcgccgctggcg ggcagttccg gcacgtcggg cccggttgcc 38760 gggtggtgcg cggtgagggcgggcaggtcc tccacggtga gtaccggcgt gccctcgggc 38820 agccgggccg cgagccgggtgtcggtgagg gtcaggcacg gcgtggcgtc ggcgacgatc 38880 ccggccaggc gccccgcggggtgctcactc tccaggggta cgaggacggc tcccgccttg 38940 agcacggcga ggaggcagaccaacaggtcg aggccgcgcg gcagcagcag cgccacccgg 39000 tcctcggggc cgatgccgcgggcgcgcagc aggtgggcca gccggccggc ccgctcgtcc 39060 agctcccggt aggtgagacggtgcccggtg gcggggggcg ggacgtcggc gacgaccgcg 39120 gtggcgtcgg gggtggccgccgcgtgggcg tggaaggcgt cgaggaggcc gtcgccgccc 39180 ggggccggcg cgtcgcggcccgaccagtgc agggccagcg tggtgtgctc ctcctcggtc 39240 aggaccggca ggtcgcgtacggcggtgtcg ccgtcggagg cgatcgcggc ggtgaggacc 39300 tggtggacgg ctcgggcgatgcgttcggcg gtgtcctggt cgaacaggtc gaaggagtac 39360 gacagccagc cggtcaggccgccgccgtgc tcggcgggcg agaactccag ggacaggtcg 39420 atgtcggacc cggtcttgggcaggttgagc gtctcggcgc gtacgccggc cagttccagg 39480 tggcgcaggg gcacgttgcgcagcgcgaac agggcgcgca ccagggccga gcggccgctg 39540 gtgcgttcgc cgccgacctcgcgcacgatc cgcgggaagg gcacgtcctg gtgggcgaag 39600 ccgtcgaggc tgctgtcgcgggtggcgcgc agcagctcgc ggaaggaggc ggcgccgtcg 39660 acgggcacgc gcagcgggagggtgttgacg aagaagccga tgagcggctc cgattcgggc 39720 cgggtgcgtc cggcgacggcggtgccgatg accaggtccc gcgcggccga ccagcggtgc 39780 agcaccacgg cgtacgcggcgagcagcacc atgaacgggg tcgccttctc ggtgcggccg 39840 agccggtcgg cccgctcgacgagctgggcg tccaggtcga tgcggacggc ggcgccggag 39900 aacgtcggcc tggccgggcggggccggtcg ccggcgagtt ccagagggag caccccgtcg 39960 agctgttcgc gccagtgggcgatctgccgt gcggcctcgt caccggccag ccatgcgtga 40020 tgggcggccg cgaagtccgggtactgcacg gcgagttcgg gcaggttcgc ggggcgcccg 40080 gtgagctggg ccgcgtacccctcggtgagt tcgtcgacga gcacgccgag cgaccagccg 40140 tccacgacga tgtggtggacgcacagggtg agcagccagg agcggccgtc gggggcctcg 40200 gggtcgtcgg tgagctgccacagcttggcg cgcagcagcg gcgcggcgga caggtcgaag 40260 gggcgccggg cgatccggtcgaggagtgcg ggcacctcgt cggcgcgggc ccggccgcgt 40320 ccgagtgcga cgcgcacctcgtcgggggtg ccgacgacct gcaccacgcc gtgttcgggg 40380 tcggtcgcga agcgggtgcgcagcgcctcg tggcgggcga cgaccgcggc gagggcccgc 40440 tccagcgctt gggtgtccaggtcgccgcgc agccgcaccg cgatcggcag caggtagttc 40500 accgcggacg ggtcgagctggtactggtag tagagctgct gctggccggg ggatgccacg 40560 tagcgctgtg cctccccggtgcgcggcaga gccgtgaccg agatgccgga gccggccctg 40620 ccgcgggcca ggtggcgggcgagcagggcg cgccgctcgg ggctgagctc tgccacggcg 40680 cgctgcaggg cggaggcgtcagcgggcagg gtctcggtca tcgctgtccc tccgcatcgg 40740 ccgcgacggg gtcgaagtcgacgatcagta cgtcgcggtg gctggggcgg gccgggtcga 40800 cgggggccag gggtgtcacgtcgtgcagca ccttgcggtc gtcgaccagg agctggtcgc 40860 cgggttcgcc gagggtgaccgcgaacagct ggttgccgtc gtcgtcgtag agggagctct 40920 cgccgccact gacgttggtgcggttgacca ggagcgaggt gacgtaggcc ccgccgtcgc 40980 ggtggcggcc ctgaggggcgggcctgccgg tctggtcggg gctggtcacg atccggaacg 41040 ggtgcagctt gacatcccagagccggacgc cttcggcccg ggagaacacg tcgccgagca 41100 ggtgcacgac ggcggccgtcagagggtcgg cgaggaacgc ctcggtgagc gggtcgaagc 41160 tgcgctgcac gccgccgttgagcgggttga gggcgctgtc ctggaagtag ggggcatggg 41220 gctgcgcggt cagctcgccggtgcggacgt cgaggcggaa gtgcccgtag cgccggtagc 41280 ggtaggtgcc cccgtccttcatgaaggtgt cgagggtgag gtcgtcccag tggcgggcga 41340 agcgtgccca gtcctgcgggccggtgccgg tgagccggcg cacctccggt gcggtcagca 41400 gatggccgcc ccatcccgacaggcctgtga cggcggcggt caccgtccgc gtctgggcgg 41460 cggccgtggt ggtctcggacacgaggtcga gcccctttcg gtgggcggcc ggggacaggg 41520 ccccggcggc ggtgcggacggatcgcatcg gcacgcgtcg cccaggtgag gccccgcgag 41580 cggtctttgg atgcaacaggcccgcacgcc ggtgcggcag tcaggacatt gcctgactgc 41640 cgcacgcgtc cgggccgaccgaccatcccg gtaagggctg cccgagcagt cggacaggga 41700 gggtgacgat gacggccgacgattcgctga cggtggtgac ctcgttcgcc cagcagagcc 41760 tgtggctgca gcatcaggtggaccccgacc gtgctgcgta caacatcact gcggcggtac 41820 ggctgcgcgg gccgctggacagcgcggcgc tggagcgggc gctgaacgcg gtggtggcgc 41880 ggcacgaggt gctgcgcacggacttcgccc tggacggcga ggacccggtg caggtggtac 41940 ggccggagcg cgaactgcgggtcccggtgc tcgacgccgg gcccgacgac ttcctgcgtt 42000 acgccggcaa ggtgctgttcgcgcccttcg acctgcgcgg cgggccgctg ctgcgcctgg 42060 agctgctgcg gctcggaccggagcaccaca tcgcgctcct ggcgctgcac cacatcgtca 42120 ccgacggggc gtcctcggcgatcctgctgc aggagctgtc ccacgcgtac gccgccgaac 42180 tggccggcgt ggagccgctgtgggaagaac tgcccatcca gtacgccgac ttcgcggtgt 42240 ggcagcgtga cgcgctcagcggtgccgccc tcgacgagct gaccgggcac tggacgcgga 42300 ccctggcggg cgccgagccgctcgcgctgc ccgccgaccg ggagcgggac gccgacgctt 42360 ccgggcacgc cgccgtgcaccacttcaccc tgcccgccgc actcacggca cgcctcgagg 42420 atcttgcccg cgccgagcgggccacgctgt tcatggtgct gctcgccgga ctcgacgcgc 42480 tgctcgcacg ctactgcgggcaggaggaca tcacggtcgt ctcccccatg gccggccgcg 42540 gccgccccga actggaggggctgatcggct acttcgtcaa tccgctgctg ctgcgcaccc 42600 aggtgtccgg cactgcgggcttcgccgcgc tggtgggccg gacccgcgac acctgcctgg 42660 acgccttcga ccgcgaggagctgcccttcg agcaggccgt cgaggtgctg cgcggacacg 42720 gcgctcctgg cgccgagagcctgcgttcgc aggtgatgct ggtgctgcag aacacgcggc 42780 cgaaccactg gcacagcgccggcatcgact tcgaactgat ccccgccgac acgtcgaccg 42840 ccaagtcgga cctcgtgctcgaggtacgcc ccggcgccga cggcggctgc acggccgccc 42900 tcgagtaccg cacgggcctgtacgacgccg cgaccgccga ccgcgtcgcc cgccacttcg 42960 ccgccatcct ggaggccgcctgcgagcggc ccgacgagcc cctgtcgcag ctgctggcaa 43020 ccaccgtccc cgacggggagatccccgcgg tgcgcagcgg cgccgcgcac ggcgaagctc 43080 ccgccgcgac cgagtacgtcgcgccccgca ccccgctcga ggaggaggtc gccgggatct 43140 ggtcggcgct cctcggcacggaacgcgtcg gcgtgcacga caacttcttc gacctgggcg 43200 gccagtcact gctcgcggtgcggctcgcgg ccaggctgcg ggacgagttc ggcgtcgaga 43260 tgacggtgcg cgacctgtatgccgacttca cgctcgaggc ggtcacctgg gccgtgctgc 43320 agcggatgac ggccgacggggaacaggcgc ccgcctgagc gaccgcagtc cgatccacac 43380 cacccagcgg acccacggattttggagaga catgttgagt gcggcggttc tgtgcttcgc 43440 gcccgccggt gcgggagcgagcttcttcca cccgtggctg gggcagcgcg agggccttac 43500 ggtgctcccg gtcgaactgcccggccggga gaagcggttc gccgagccgg agtgcaccga 43560 gctcggcgcc ctgatggacgcggtcctgcc cgccctgcgg tccgcgacgg ccgggttcga 43620 caaggtcgcc gtcctcggacacagcttcgg cgcgctgctc gcctacgagg cgacgcgcgc 43680 gctcgccgcg gagcgggacgggctgacgct ggtggcctcc ggcgcgacgt ggccgggcac 43740 actgcgccgc gggcggatcaccggcctggc cgacgacgag ttcatcgccg gtatccacac 43800 gatcgccgga taccgccacccggccctgga cgagccggag ttgcgcgatc tggtgctgcc 43860 gccgctgcgc accgacgtcgcgatgcacga gaactacgtg tacgtcgagc gtgagcccct 43920 cgacgtgccg gtcctcgcggtacgcggcga cgccgaccgg ctggtgtcgg ccgacgacct 43980 caccgagtgg gagcgggtcaccaccggcac cgtgaccgtc gcggagctca ccggtgccca 44040 catgtatctg gtggaggagtggcccgcgct gctcgacctg ctcgccgggc acctcctcgg 44100 cgccgggacg ccgtcatgagcggacggctc acgggccgga ccgcgatcgt caccggcgcc 44160 gggcggggcg tcgggcgtgcctgcgccacg gcgttcgccg cgcagggcgc cgacctggtc 44220 ctggtcgaca tcgccgccgacctcccgcac gtcccctacc ccgcggccac cccgagccag 44280 ctcgatcaca ccgcccggctgtgccgcgag cagggggccg ccgtgctcac cgcacgcgcg 44340 gacgtgcggg acgcggcggcctgcgagcgg gtggtggccg atgccgtgga ccgcttcggt 44400 tcgctggacg tgctggtcaacaacgcgggg atcgcgggtc cttcgggccg gatcgtgcac 44460 gaggtcaccg aggacgagtgggcggtgatg atcgacgtca acctcaacgg cgcctggcgg 44520 atgctgaagg ccgcgggggcgtcgatggtg gcggcccgct ccggctccat cgtcaacatc 44580 gcctccaccg ccggactggtgggataccgc aacttcgccg gctacgtggc gtccaagcac 44640 ggcctggtcg gcctgacgaaggccgccgcc ctcgactacg cgccctaccg ggtgcgggtg 44700 aacgcggtct gccccggctcggtgcgcgac ggcgaggcgt gggagggccg gatgctggtg 44760 gagatcggcc gcagcatcgggatcgaaccg gccgaccacg aagccgagtt catcacgcag 44820 atgcccatga acacgctggtggaggcggac gatgtggcgg gggccgccct gtggctcgcc 44880 tccgaggagt cccgtcactccaccggcggt gtgatcaccg tcgacgccgg ctacagcgcc 44940 cgttgacccg gaccggtcaggagacgcccg tgtcgtccaa ctccccttcg gtcgccgccc 45000 ggcacgccgc cgtgtacgcggtcgcccccg gatactcggc cgaggactgc cgtgccgcgc 45060 tcgccggcgc cccgtggcccggccccggca ggttgcgcct gtgggccgag ccggtgcccg 45120 ccccctcgga cagccccgctgccctgcgcc gccgcacccg cgagctgcac ggccgtcacg 45180 ggggcgaagg tgtccggctcaccctgctcg aatacgccga cggggtacgg gatctggtgg 45240 ccgtgaccgg ccccggcgacgacggcgccg accgcctgga ccgcgccgcg ctggccgccc 45300 tcggcccggc gcggcccctggccggctcgg gatccccttc ggaccccggg tgccgggcgg 45360 tgccggtccc ggggctcggcgcgggcgggg ccatggccac ggcctgggac ctgacggcgg 45420 ctctgctcct gatcgccgccaggacgcggg cgcaggacga gccggtcctg gacgtcgccg 45480 tccacaccac cgacggggtgcgccgcgtct cgctccgcgt ggccgcgttg gacgaggacc 45540 agccggtgtc cgcctaccgggcctgggtgc gcgagcaggc ggagcaaacg agtgcccagg 45600 acggtggcgg cgggccgcggctcgccctcg atgtggacct gcggccgcgg gcggtgcccg 45660 cggtgcggcg gctgcccgcgctgcacgccc gccacccggt caccgtgcgg gtcaccgcct 45720 ccgacgcgga cggcaccccgcaggtgcact gctggaccct gcccggaacg cccgcggacc 45780 agctgccgac agactttgaacagcaacttg ccgtagtggc acaccagctg ctgtgcgggg 45840 accagaatct cccgctggccgacgtcgcgt tgcacgagcc ggccgagcga gcccgcatcc 45900 tggatctcgg ccgcaccccgcgcgccgcgg acagcgggcc gcgccgcatc gaccagctgg 45960 tccgggaccg cgccgagcggaccccggacg ccgtcgccct gagagacccg cagggcgagc 46020 acacctggac gtacggtgaactcgtcgaca ggagcgaccg gttcgcggcc gctctgcgag 46080 ggctcggtgt gcgcccgggcgaccgcgtag gggtgtgcct ggaccgttcg gcacagctgg 46140 tgtcggtgct cctcgcggtgatgacggcgg gcgccgccta cgtaccgctg gaccccacct 46200 atccggccga ccgcctggcctacaccgccg acgacgcggg cctgtccctg gtggtggtcg 46260 aggacggcgg caaggacgacggcaacgcct tcgccgacca cgccaccgtg accctgccac 46320 gcctgcggga actcgccgccggacagggcg cgtgggagga gcccggcacg gtggggccgg 46380 acgatccggc gtacatcatctacacgtcgg gctccaccgg gcgccccaag ggcgtcgtcg 46440 tgccacaccg caacgtggggcgcctcctgg atgccacggc cgacgacttc cgcctgggcc 46500 cgcaggacgt gtggacgtggttccactcgg cggcgttcga cttctcggtc tgggagatct 46560 ggggcgctct gggcaccggcgggcgcctgg tcgtcgtgcc gtactggacc tgccgctccc 46620 ccgaggactt ccgcgcgctgctcctcgacg aacgggtgac ggtgctcaac cagaccccgt 46680 cggccttctc ccggctgctgcccctggagc gcgcggcgcc gacgccgctc gccctgcggc 46740 tcgtcgtctt cggcggggagccgctcgacg cccgcgccct gctcccgtgg ttcgacacgc 46800 atccggagtc cgcgtgccggatggtcaaca tgttcggcat cacggagacc accgtgcacg 46860 tgacggccca gaccgtcacccgcgccgacg ccctcgcctc ctcgcagtcg gtgggccgcg 46920 cgctgcccgg ctggagcgtgcgggtcctcg acacacgcgg caggctggtg cagcccggct 46980 gcgtgggcga gatagcggtcggcggggacg gcctcgccct ggagtacctg ggacggcccg 47040 agctgaccgc cgagcggttcgtacccgacc cggacggcga aggccgcctc tacctcagcg 47100 gcgacaaggg ccggcagctgcccgacggcc gcctggagca tctgggccgc ctcgacagcc 47160 aggtcaagct gcgcggtcaccgcatcgaac tcgacgagat ccgctcggtg ctgctcaccc 47220 acccctcggt gcgtgccgcggcggtcgtac tgacccgccc cacggacgcg aacggcgagg 47280 acgccaccct cgacgcatacgccgtactgg acggggccga cgcccgcgag gtccgccggc 47340 acgccgcccg gctgctgcccgagtacatgg tgcccgccac cgtcaccccg ctcgccgagc 47400 tgccactcac cgtgaacgggaaggtggacg tggccgcgct gcccgcgccc cgggccgccg 47460 gcggtgaact cccttgtgtggcggacgagt ccgcacaggg cacgctggcc gccgtgctcg 47520 ccgcctggca caccgcgttcggcgaacagg tcgcggccgg cgacgacttc ttcgagctgg 47580 gcggcaactc actgcgcgcactgcgcgtcg tgcacctgct gcgggacgcg ggcctccgtg 47640 tcgacgtgcg cgacgtgtaccggctgcgca ccccgtcggc gctggccgag gccgcctcca 47700 cggcgcccgc cgacgcggcgaggacggcct gacccggccg tgtaagggct ttgcccgact 47760 gtccgggacg ccaccggcggcgggatgttc gagacctgac accgattcga ggagtgttca 47820 tgagcaaggt cgcggtcctcaccaatgacc ttcagtacga cctggtgaac aagaaccagg 47880 agcgcatcga cgccgtggcggaggccaccc ccagcttctc gggcttcctc aaggacatgc 47940 gccagcgcgg ccaccacgtgctgcacctgc agctgatcaa caaggcggac gacccggtcg 48000 ccgagcggta cgagggcggccgactgcccg cgcagaaggg cacccagggc gccgagatcc 48060 tgcccgtctt cctggacgacaccgacatcc tcgtcgagaa gaacaaggac agcggcttct 48120 acgagacgga cctgcacgagcggctgcagg ccctcggcgt cgagaccgtc atcgtgaccg 48180 gcatgcaggg ccagatctgcgtgcagacca cggccgccga cgcgttcttc cgcggctaca 48240 acgtctgggt ccccagcgactgcgtcgtct cggcccagga gcacgacaag gtgcgcgccc 48300 tggactggct ggacggctactgcgcgaccg tggcgccgtc cgatgagatc gtccgcgtcc 48360 tcgacgccga cggcgacctgccgcgcaagg tgttccgcac cccctgagcc ctcggtccgc 48420 gaacccgcga ccggcctcacaggcgggcgg cgcccctccc cgcgccgccc cctcatcacc 48480 ccgcgtcacc ccgcttcagccgccccctca tccgcgtacg tcttcgaccc caccgggagt 48540 cacgttgccg acggccacgacgtcctttct ggaccagaaa atcgtccttg accacgggac 48600 cggggcccgg ctcagccgggaactcgtcga gttcgtcgtg gaggtgctcg gggacgtcta 48660 cctcggcgag atggaggacagcgccatcct cccgccgctg gacggcgctc tcgccatgac 48720 caccgactcc ttcgtggtggacccgccgtt cttcggcaac ggcgacatcg gcaagatcgc 48780 ggtgtgcggc accgtcaacgacctcgcggt ggccggcgcc gagccccgct acctcacgct 48840 ggcgatgatc ctggagagcgggttcccgct ctccgggctg caccgcgccc tggagtccgt 48900 ccgggacacc gcgcgcgaggcgggcatcaa gatcgtcgcc ggtgacacca aggtcgtcgg 48960 ggagggcgag gccgacggcatcttcctcaa caccaccggc gtcggcgtct tcacctcccc 49020 gcccaagcgc atgaaggacgtggccgtagg cgacaagatc ctgctcagcg gccccatcgg 49080 cgaccacacc atccacctcctctcggtgcg cgagggcctc ggcttcgaga agaacgtgct 49140 cagtgactgc gccccgctgaacaacatgat cgacacggtc cgcggcgcgg tgcccgagtc 49200 cgcggtgcgc tccatgcgcgacgtgacgcg cggcggcctc aacgcggtgc tgcacgagta 49260 cgcctccgcc ctcggtcacgccgtacgggt cgaggaggag gcgctgccga tcagcttcga 49320 gacggcgatg gccgcggacatgctcggcat caacccgatc aactgcgcca acgagggctg 49380 tctggcgctg ttcgtggcgcccgagcacgc ggacgaggtc ctgcgcgtgc tgcgcagcca 49440 cccctacggc aaggacgccgtcatcgtcgg cgaggtcacc gcacgcccgg agccggtcgt 49500 ggagatggcc ggcaaggacggccggatcca ggagatcgag gaactgcagg gcgccgagct 49560 tccccgcctg tgctgatccgaggacccgag gaggagtcat ggcagagacc gagcagtggc 49620 tgatccgggt gaccggggtggtgcagggcg tgggctaccg gcccttcgtg cacaccctgg 49680 ccaccggcct gggcctgagaggctgggtcc tcaacgacac gctgggcgtc cagaccgagg 49740 tgtccggcgg ggccggcgaactcgcggaat tcgcgggggc gctcaaggac cgcgccccgc 49800 tgctcgcccg cgtcgaagaggtgcgggtgg agtcgcggac gccccgggcg ggcggtgacc 49860 tgccgacggg cttcgagatccgggagagcc ggcgcaccgg gcgggcgaac accatcgtcg 49920 ccccggactc gcacgtgtgcgacgactgcc tgcgcgaggt cctcgacccc accgaccgcc 49980 ggttccgcta cccgttcaccaactgcaccc actgcgggcc ccgttactcc ctgatcaagg 50040 gcctgcccta cgaccgtgagaagaccacga tggccgtctt cgcgatgtgc gaggactgcc 50100 gcgccgagta tgtgaaccccgcggaccgcc gctaccacgc gcagcccacc gcctgcccgc 50160 ggtgcggccc gaaggtggccctgtccgcgc ccgacggggt ggtggccgag gccgatgagg 50220 cgctgacgcg cacgatcagggcgctcggcg acggccagat cgtcgccgtc aagagcgtcg 50280 gcgggttcca cctcgcggtgaacgcgcgtg acgcggcggc cgtggcccgg ctgcggcgcc 50340 gcaagaagcg ggactccaagccgttcgcgg tgatggccgc gggcctcgac gagacggccg 50400 caatcgccca catcacggacgccgagcggg agttgctgcg ttccccggcc cgccccatcg 50460 tgctgctgcg caagctgccgggcgcactgc ccgaggaggt cgccccgcgc aacccgaacc 50520 tcggtgtgat gctgccgtcggctccgctgc accaactgct tctcgaccgg ccggagtcgg 50580 acgtcctcgt gatgaccagcgggaacatct ccggccagcc gatcgcgtac cgcaacgagg 50640 acgcgctggc cgagctgttcgaggtcgccg acctgatcct gcaccacgac cgggacatcc 50700 acgtccgcgt ggacgactcggtggtacgca gctccgtgca cccggccctg gacgagccga 50760 tcatcacgtt cctgcggcgctcgcgcggct acgcccccta cccggtgaag gtcgacgaca 50820 tcgccggacc ggtcatcgcgtacggcgcgg agctcaagac cacggtcgcg ctcggcgacg 50880 gcaacgaggt ctacgtcagccagcacatcg gcgacctgaa gaacgacgag acctacgcct 50940 cgcaccgcga ggccgcggcccacctcgccg aactgtacga gctgaagccc cggcacgccg 51000 cctgcgacct gcaccccgccttcaaggccc gggtgctggc cgggaccgac cggcccgacg 51060 acgtcgtcga ggtccagcaccaccatgccc acatggcgtc ctgcatggcc gagaaccggc 51120 tcagcggacc caccatcggcgtcgtcttcg acggcgccgg ctacggcgag gacggcacca 51180 tctggggcgg cgagttcctcgtcggcgact tcgcctcggt gcgccgggcg gcccatctgc 51240 ggccggtccc gctgctcggcggcgacaagg cggtggccga gccggtgcgc accggatacg 51300 cgctcgcgct gcaggcactcggctcccccg agcgggccgt cgccgcgttc cccgcgctgg 51360 agatcctcga cgaccgcaagcgcaccgtgt tcgcgaccat ggccgaacgc ggcatccagg 51420 ccccgccggt gtccagcatgggacggctct tcgacgcggt ggccgccctg ctcggcgtgt 51480 gcgcctacgc cgagtacgaggcacacggcc cgatcgaact ggaggggctg ctgggccgcg 51540 acctcgccct gaccgcgccctacgccttcg gactccagca ggagggcgac accgagctcg 51600 tcgacccgag gccgctcgtgcgggcgatcg ccgcggacct ggcacgcggc accggcgtca 51660 ccgacatcag ccggcgcttccactccacgg tggtcgacat ggtcgtcaag cggtgcagcg 51720 cgctgcgcga acgcaccggcatccgccaag tggtgctctc cggcggcgtc ttcctcaacg 51780 agttccttca ggtcaactgcctcgtggagc tgcgcagggc gggcttcgac gcccatgcgc 51840 accgcctggt gcccaccaacgacggcggta tcgcgctcgg ccaggtgatg gtcgcgaacg 51900 cacgcctgcg tgccagcccggacagcacag acagcccggc caccccgaac cccggcacga 51960 acgaatgccg ctgacccgctccaccgacga cggaggcaca gcccgatgcg gaccgcagat 52020 ctgacgctgg acaccgtcgtgcgcctgctg agtgagaacg aggcgagcgc acggcagacc 52080 ctgagcatgg tgccgtcggagacctccatg tcggggctgg ccaagttgcc catgcttctc 52140 gacccgtacc accgctacttcttcaacgag ggcgacgacc ccgaccgctg gcacttccgt 52200 ggcgcgcagc ggctgcgcgacctggagatg gagctgacca tcccgctcct gcaggagctg 52260 ggccgggcct cgtacgcctcggtgcggccg ctgagcggtc tgaacgggat gacgctggtg 52320 ctcggcgcgc tcggcggcgaaccgggttcc acggtcgtca ccgtctcccc ggagcaggga 52380 ggccactacg cgacgccacaggtggccggc cggctcggcc tccatgcgga gttcctgcgc 52440 ggccccgacc cgcactccctcgacctcgag cacgcggccg agctcctcgc ccgggtacga 52500 ccctcgctgg tgtacgtggaccagtcgcac tgcctgttcc ccgtcgatgt gaagtccttg 52560 gtggagacgg tgcgggaggcctcgcccggc acgctggtgc acgtggacgc gagtcactgg 52620 ctcggcctgg tactcggggacgcgttcccc aacccgctgg accagggcgc ggacagctgg 52680 ggcggctcca cccacaagaccttccccggc ccgcagaagg cggtcgtgct gacgcgggac 52740 ccacaggtgg agcagctgatccgcgacgcc caggacttcc tcatcagcaa tcaccacttc 52800 gccgcgacca tcgcgctgggcatatcgctc ctcgagttcc gcgagttcgg ccccgcctac 52860 acccgggcgg tcctggagcacaccggccgc ttcggcagac tcctgaccga gcggggcctg 52920 accgtggtcg ccgccgaccgcggctactcc gcggggcacc agctgtggct ggacacggag 52980 gccgacggca tcgcgccgaaggacgcggcc gcccggctct ccgccgccag cctcaaggtc 53040 aacttcatgg ccgggctgccgggcttcacc gggcagggcg tgcgcatcgg cctcaacgag 53100 gccacgtacc aagggctttcgggtgacgac atcgacgagc tcgccgacat cttcgtggcg 53160 gccgtgcgcg acaccgaaccggccgccctg ctcgccgagc gcacggccgc gctgcgcacc 53220 cggaccccgt tcggcgcttgtgcggccgat ggcagtccgc tgctcgccac ggctctggac 53280 ctgtgcgagg gcgccctgcggcgcaccggc gcggccgccg ggcgtcccct gacagaggcg 53340 tcccggtgag ccggccggtcatcggcatcg cctcgtaccg cgaccgcgcg cgctggaaca 53400 tctgggacac ggacgcgacggtgctccagc aggggtacgt cgacggggtg cgccgggcgg 53460 gcggccgtgc cgtcgtgctgccgcccgacg acaccgacgc ggacgtggtg gcccgtctcg 53520 acggactcct gctgcccggcggtgccgacg tcgacccggc gcgctacggt gccgcccgcc 53580 atccgcgcac ggacactccgtcggccgacc gggacgccgg ggagctgctg ctgctcgagg 53640 cggcgctcgc cgcgggactgcccgtcctcg gggtgtgccg cgggctccag ctgctcgccc 53700 tcgcctacgg cggcaccctgcaccagcatc tgcccgatct ggtcgggcac agcggccact 53760 gcccggccga gggcgtcttcggccggcacc ccgtgcacct ggtgcccggc agccgggccg 53820 ccgcgctgta cggggagctggccgaggtca actcgcacca ccatcaggcc gtgctcgatc 53880 cgggcggact gcacgtgaccggacgtggcg aggacggcgt ggtggaggcg gccgaggacc 53940 cgtccaggcc gttcgtcctcggcgtccagt ggcaccccga agtctccgac gaccggaagc 54000 tgttcgcggc gttcgtcgcggcctgcacca cccggtcgct gcacacagcg gcatccgtct 54060 gaccgcgccg gcgccgtgacgccggtgcgg tggcacgtgc cctcgcccga ccgggccccg 54120 tacacctcac cgtaggactttgccaggagg gcaaccgtgc acactcacgt ccgtcccgtc 54180 ggcggcacac tggtcgacatcctgcgcgac cgggcggacg ccactccgca ccgcaccgcc 54240 ctggaattca tccgcgacgacggcagcgaa cacgccgtcg gctacgcggc gttggaccgc 54300 agggtgcgcg ccgtcgccgccgcgctcacc gagcgcggcc tgcgtggcga gcgcgtcctg 54360 ctgctgttcc cgccgggcgagaactatgtg acggccttcc tcggctgtct gtacgcgggc 54420 gctgtcgccg tgcccgtgtatgtgccgacc ggcaagcgcg gcctgtcggc ggtccttgcg 54480 accggggcgg acgcgggcgccgtgctcgcg ctgagcagcc gcgaggtgac ggacgcgatc 54540 acggcctcgt acccggagctgaccacgtcg gggctgatcc agtggctggt caccgaggac 54600 atcgcggacg cggcggccgacgactggccg ggcaccggcc ccgcgcccgc cgacctggcc 54660 ttcctgcagt acacgtccgggtccaccggg acgcccaagg gcgtgatggt gcggcacgac 54720 aacctggtgc acaactcggcgtccatcagc accgccctgg gcgtcggtcc ggacagccgg 54780 ggcgtgagct ggctgccgccgtatcacgac atgggcctga tcggcggcat cctgcagccc 54840 ttgtacgcgg gtttcccgtgcaccctgatc tccccgatgg cgttcgtacg cagcccgtac 54900 cgctggctcg acgccatcagcaggcaccgc gcgacggtgt cggcggcgcc cgacttcgcg 54960 tacggcgagt gtgtgcgccgcattcccgag gacaagcgcg ccgagctcga tctgagcagc 55020 tggcagcacg ccatggtgggtgccgaacct gtgcggcccg cgaccctgga cgccttcgcg 55080 cgcgccttcg cggacagcgggttcgaccgg tcggccttcc acccctgcta cgggctcgcc 55140 gaggcgacgc tgttcgtcaccggcggcgcg cccgagcggg gcgaaccccg ggtgctgcgc 55200 gccgaccgtc gcgaactggagctgggccag gccgtgtcgg ccatcgcggg caaggagaac 55260 aaggagaacg ctgctgtgctgacgggctgc ggccggtcgc gcagcgagga cgtcgtcgtg 55320 gtggtggatc cggccgagggccgtgcgtgc ccgcccggcg ggaccggcga ggtgtgggtg 55380 agcggcccga ccgtcaccgcgggctactgg ggccggcccg agctgaccga cgaggtcttc 55440 cgcgccaccc tggagtcccacccggggcgg cacttcctgc gcaccggtga cctcgggttc 55500 gccctgggcg acgagctgttcgtgaccggc cgcgccaagg acctcctggt ggtgcgcggc 55560 cgcaaccact acccgcaggacatcgagcag agcgccgagc acgcccaccc gctgctgcag 55620 ccgacccggg cggcggtgttctcgcacgac gacgggaccg aggagcgggc cgtcctggtg 55680 cacgaggtgg tccgcggcttcgaccccgaa cgcgcccccg aggtgatcac ggcggtccgc 55740 aaggcggtga cggccgaacacgggctctcc ctgcacgagg tggtgctggt gcggcccgga 55800 gcgattccgc gtaccacgagcggcaaggtg cgccgcagca cctgccgcga gcggtggctg 55860 gacggcactc tgcagccgatcgcgaccggc ggcgcggaca ccgcgcccga cgccgggggc 55920 ggtgacatgc ccccggtgtcgagggcggtc gccgaggtcg tcgccctgga actcgacgtg 55980 cccgccgaac agctgccgcccggtgtgccg ttggtggtcc tcgacctcga ctcgctgcgg 56040 gccgtgcggc tcgccgaagcgatgcgtgag cgcttcgggc tccgggtgtc ccccgccgag 56100 ctcctcgacg ggctgacgcttgcgggcctg agcgccctcg tcgacgccgt cgacgacgtg 56160 gccgaggcac acgtcggtgccgacgacacc gaggccgccg acgcgcccgc gagccgggcc 56220 caggagtgga tgtggctcctcgacgccatg ggcggcggcg cggcgtacca cgtggcgggc 56280 ggcatcgacc tgcacggccctgtcgacccg gagttgctgc gcgcgagcct ggaggacctg 56340 gtggcgcgcc acccggcgctgcacagcggt ttcacctccg accccgatgg caccctgcgg 56400 cgcacgccgc tcgcccctcgctccttcgag ctgccggtgg tggagccgag cgcggaccgg 56460 gacgcccgcg cagtcgcggccgaactcgcc cgggagccgt tcgacctggc ggcggggccg 56520 ttgctgcgcg cggtcctcgtgcgggccggc gagcacgagt ggacgctcgc ggtggccgcc 56580 caccacatcg cggtggacggctggtcactg ggtctgctgc tgcgcgaact cgggatctgc 56640 taccgcgacc ggctcgcggggcgcagcccg tcgcgcaccg tcgtcacggt tcccgcgccg 56700 aagagcgcgg gcgacgacgcggaggccgcc gcgttctggc aggagaccct ggccggggcg 56760 caggcggtgt cgctgccgctggacccggcc ccgcacccga gccccttcaa gggctccgcc 56820 ctgcccttca ccctgcccgcgccggtggtg gcccggctca aacgctatgg ggccaagcgc 56880 cgttcgaccc tgttcatggtgatcctcacc ggcctgtccg ctgtgctgag ccggtggacc 56940 ggccagcagg acctggtggtcggcgccgtt acggccggcc gccaccgtcc cggcagcggt 57000 gacctggtcg ggctgttcgtcaacaccctg ccgttgcgcg tcgaggcctc cggtgacccc 57060 gccttcggcg agctgctgtcccgggtccgc tcctcgtgcc tggccgccta ccggcaccag 57120 gacctccccc tcgaggagatcgtccggctg accggcgcgg accgcgccga cggacgggcc 57180 ccactggtgc gcgtggcgctcgccctgcag aacctgccgg tggcaccctg gcagtccggc 57240 gaggtgcgcg ccgagcccttcgagctccct tctcccggcg cgcagttcga gctgtcgctg 57300 catctgacgg aggagagcgacggcagcctg acgggccacg ccgtccacgc ctcggacctg 57360 atctccgcac cgagcgcgcgccgcatcctt caggccctga cgctggccct cgaagccgtg 57420 ccggggccgc cgggggcggcggcgacagat cttccgatgc tcgccccgca cgaactcgac 57480 cgggtggtga gcgagttcagcggctcggac acgccgctcg acgacggcct ggtgcacgcc 57540 ggtttcgagc ggcaggcccggctcaccccg aacgcaccgg ccgtcgtgtg ggacggcggc 57600 acgctgtcgt acgcacagctggacgcccgg gcgaaccggc tggccaggcg gctgcgctcc 57660 ctgggcgcgg cccccgatcaggtggtggcg gtgcacctgc cgcgctcccc cgagctggtc 57720 gtcgcgctcc ttgcggtcgtcaaagcgggc ggtgcctatc tgccgctcga cccggaccac 57780 ccgcgcgccc ggctcgccctgcaactcgcc gacgtgaccc cgtcgctgct ggtgacgtcg 57840 cgggatgccg cgcacggcagagcgggcgaa ctcgcggcag gctcgcccgg gctgacgctc 57900 gtcgagatcg acgacccggccctggccgaa ctgccggacg gccccctcga cgaggtcgcg 57960 cggccggaca acctgctgtacgtcctgcac acctctggca cgacgggcac ccccaagggg 58020 gtgatgaacc ggcatgccggcgtggccaac cggatggcct ggatgcagga gaagtatccg 58080 ctcgtggccg gggagcgcgtgctgcacaag acgcccgtcg ggttcgacgt gtccggctgg 58140 gagatctggt ggccgctcggccacggcgcg acgatcgtgc tcgcccggcc cggcggccac 58200 caggatcccg ggtatctggtgcggtccgtc gtcgaccacg cggtgacgac ctgtcacttc 58260 gtgccgtcca tgctgcgcgtgttcctcgac gaaaccacgg ccgcgacggc cggggggacg 58320 ctgcgccggg tcgtgtgcagcggtgaggca ctgccgccgg acgtggccgc gcggttccac 58380 cgtctgctgc cgggcgtggagctgcacaac ctgtacgggc cgaccgaggc cgccatcgac 58440 gtcaccgcgg agcaggtcgagcccggcgcg accgaccggc cccggctgcc gatcggacgc 58500 ccggtgcccg gcgtacgcctgtacgtgctg gacggacggg gcaatcccgc gcccgtcggc 58560 gtacccggcg aactgcacatcggcggcccc caggtggcgc gcggctactt cgggcggccg 58620 tcgctgaccg cggccgccttcgtccccgac ccgttcggct ccggggggcg gctctaccgc 58680 accggcgacc gggcgtgctggaccgaggac ggcaccctcg actacctcgg ccgcatcgac 58740 caccaggtca agatccgcggccagcgggtg gagcccggcg agtcggaggc ggtgctggcc 58800 aggcacccgg ccgtcggcgacgccgtggtc gtgccccgcc ggcaaggcgg cgagacgtat 58860 ctggccgcct acctggtggccgtggccggc gccgagcagc cgggcccggc acacctgcgg 58920 gagcacctct ctctcagcctgccaccggcg atgatccctt cggcgtacac atggctggac 58980 gcgctgccgg tgggcgccaacggaaagctg gaccgcgacg cgctgccgga gccggaccag 59040 aacctcgccg gcggcgagcacgtggcgccg cgcgacgcgg ccgagcagcg ggtggccgag 59100 gtgtggtgcg aggtcctgggcctcgaggag gtctcggtga cagccgactt cttcgcgctc 59160 ggcgggcact cgctgcaggcgacccggatc gcgctgcggc tgcgcgaggc gttcggggcg 59220 gagttctcgg tcgccgacctgctgaccgga gtgcccaccg tggagcagac ggcacggctt 59280 ttgacgcagc gtcaggtggcggagagcgac ccggaggagc tggcccggct cctcgaccag 59340 ttgtccgggc tgtccgacgacgaagtggcc gacctgctgc agcagcaagg ctgacggtgg 59400 ggaaaggaga cgccggcccatggccgtgac actcaaggac gacttcgccg agcgggcccg 59460 cctggaggcg cggctgctcgcccggcgcag aggcggagcg gcgctgccgc tgctcgaccg 59520 cccggccggg gccgagtggcacttcgccgc ctcgctgggc caggaaggca tgtggccctc 59580 cctggcgacg acagcgcgccagcccctgat cgtgggcggg ctcagggtgt cggggccgct 59640 ggacaccggg gcgctgcgcgaggcgtggca ggctctcgcc gagcggcacg agacgctgcg 59700 cagcgcctat cggagggtcgacggcgcgct gacgcaggtg gtcccggtcc gcgcgaggcc 59760 ggtcatggac gtggccctggccgacccggc ggcggccgcc tcggtcgcgg acgacgagct 59820 tgcccgcccc ttcgacttcggccggggccc cttggcccgg ctgcgcgtgc tgcgccgcgc 59880 ctccggtgaa catctcgtggtggtgtgcct gcaccacctg gtcggcgacg cccgggccct 59940 cgaggtggtc accaccgagctcaccgcgct ctatgccggc gccgtcacgg gctcgcccgt 60000 acagctgccc gaactcccggcccagtacgc cgacttcgcg gcctggcacc ggtcgcgcgt 60060 cgaaggtgag cacggcggacggactgtcgc gtactggctg gaccgtctgg ccggcgccga 60120 acccgcggcg ctgccctgcgaccggtccca ggacccctcg gacatcgcgg gtcacacccg 60180 gtccttcccg ctgccgccggagctcttcgc ccggctgcgc gaggtggcgg cccggcaccg 60240 caccactttc tacgtggtcgggctcgcggc cttccagatc ctgctcgcgc gctggtcggg 60300 cgagcgggac atctgcgtgcgggcacccgt ctcctaccgc gaccgcagcg aggtgcacgg 60360 cctggtcggc gacttctccaacgacgtggt ggtgcgcgcc gacctgtcgg gacacgtgac 60420 gctggccgag gccgtggagcaggtgcgggc gagcacggcc gacgacttcg cccaccacga 60480 cgtaccgccc cacgtggtggcctcgcgcat ggccgatccc accctgttgg agcggctgtt 60540 ccacgtgcag ttcaccgcggaacgcgagca cgacctcggc gccgcccggc agctgggccc 60600 gatacgcgcc gagccgttctcccccacggc cgaatccgtc gcgcgcccgc tcagcgtccg 60660 cctgcgccac gacgaccacggcggccgcgt catcgtcgcc taccgctcgg gcctcttctc 60720 cgacgagcgt gtacgggcactccagcgcga ctatttcgcc ctgttcgagg agatcgcggc 60780 acatcctggg cagccggcggtgagttgacc cgatgctcac tcctcggcag ccgatcgggc 60840 cgggtgttca agagtgagcggagttcggcg ggcggaaggg tcacgctccc agtgcgagga 60900 ggccctcatc cgaggtgcccggttgctgtc gcggaggagt gggactcatg tcggctacga 60960 ggcgggtgca catctaccctttcgagggag aggtcgacgg ccttgagatc cacccgaagt 61020 tcgcggaact gagggagaccgacccgctcg ccagagtgcg cctcccgtac ggcggcgagg 61080 ggtggatggt cacgcggtacgacgacgtgc gcgccgccaa ctccgatccc cggttcagcc 61140 gcgcgcagat cggtgaggacaccccccgga ccacaccgct ggcccgccgc agcgacacca 61200 tcctcagcct cgacccgccggagcacacac ggctgcgccg cctgctgtcc aaggccttca 61260 ccgcgcggcg catgggcgcgatgcagtcct ggctcgagga actgttcgcc gggctcctcg 61320 acggcgtgga gcgcacgggacatccggccg acatcgtgcg ggacctggcc cagccgttca 61380 ccatcgccgt gatctgccggctgctcgggg tgccgtacga ggaccggggc aggttccagc 61440 actggtccga ggtcatcatgtccaccacgg cgtacagcaa agaggaggcg gtgtcggccg 61500 acgcctcgat ccgcgcgtacctcgccgacc tggtgtcggc gcgccgggcc gcgccccacg 61560 acgacctcct gggcgtgctggtgtccgccc gggacgacga cgaccgcctc accgaggacg 61620 agctcatcac gttcggggtgacgctgctcg tggccgggca cgagacctcg gcccatcaac 61680 tgggcaacat ggtctacgcgttgctcaccc acgaggacca gctgtccctg ctgcgggagc 61740 agcccgagct gctgccccgcgcggtggagg agctgctgcg gttcgtgccc ctgggcaacg 61800 gtgtcggcaa cgcccggatcgccctggagg acgtcgagct gagcggtggc acggtgcgcg 61860 cgggtgaggg tgtcgtggccgccgcggtga acgccaaccg ggacccccgg gcgttcgacg 61920 acccggaccg cctcgacatcacccgggaga agaaccccca tctggcgttc ggtcacgggg 61980 cgcactactg cctcggtgcccagctcgccc gcatggagct gcgggtggcg atcggcgggc 62040 tgctggagcg cttcccggggctgcggctcg cggtgcccgc ggaccaggtc gagtggaaga 62100 ccggcggact gttccggggcccgcagcggc tgccgatcgc atggtgaggg gaaccggcat 62160 ggcacgggag cagaacacacgctggcggat cgaggccgac cacggcgcgt gcatgggcac 62220 cggcatctgc gccggtgtcgcaccggacgt cttccaggtg gtcgacgggc ggtccgtacc 62280 gccggccgga gaggtgccggcggacgagaa cgtgctggac gcggtcgagt cctgccccat 62340 ggaggcgctg cgggtgcgggacgcggtgag cggcgaggtg ctcgacctgt cgtgacggtg 62400 cgggcctgcg cggcgcttctcaagtggctc tccagcggaa ttcgagtggg gtgtggcagg 62460 aaaggactac ggtcggacgggtatgggcac cgcccggctg gagacgggtc ccgcggtacg 62520 ccgggaattc gcccacgtgtgccaatcccc ttgcgccgca atccagttgc acgacttgcg 62580 ccgacttgac acgattcatggggacatatg agtatccctt tggattcagg aattccgtat 62640 tccggtccgg tttttcgcacattgcgacaa ggagagcccg aatgaagttc gcgatcgtca 62700 tcatcgagaa cgccgagtcccggcgtcaga tccaggagga ccgcgcgacc taccgcaagg 62760 cgatcgaggg ctggatgggccagcaggccc aggccggtgc cctcgtgggc ggcgaggcgt 62820 tcgagaccga gtccctcgccccggtgacgg tgcgccggga ggccgacggc acccgtaagg 62880 ccgtcgaggg tccgttcgcgggcgccgacg agacgctggg cggttacatc ctcgtcgagg 62940 ccgccgaccg cgacgccgccgtggagatcg ccaagtcgtg gccgaacccc gagaccctcg 63000 aggtacggcc gctgtgggtggccgagtaac cagccggtcc caccggctcc gacccgcgtc 63060 cgggcacgcc cggccgcgggttcaggcctg taggcgctat ggagaggcac tcgccatgac 63120 ggacggcgcg atacacacacacgcggcggg agacggcgcg ggccggctgc ggccccttct 63180 ggacgccgtg cacgacacggcctgcgaggt acgggcccgc cgccccgact acttcgagca 63240 ccggccgctg cgactgcccgcggaactgct cgcgcaccac ggcaccaccc ccgccgcact 63300 cgcccaactc atcggtgattcctatgagtt gacggtcgaa ccggtcaccg accccgcgga 63360 ctgcacctcc gtggaccgtctcgcggccgc ccgcgcacag cacgccggca ccacgcccgg 63420 cgacagcggc gcccagcgcgcgccgcagcc cggcgaactg gcggccgtcg cccgcgagca 63480 gagcgagctg cgccgcgccgccgagcggga cgccgcgcgg atcggtgcgg cggcgccgtt 63540 gcgggtcggg gtgctcggcggcgtgctgcc gtacaccgcg gcgggtgtgc gcgaggaggg 63600 cgagccgccc gtcgtcctggtgaacgcgct cggcatgggg accgcgctct ggcaccccct 63660 gatggcggct ctcgcacccacgcgccgcgt gctgacctgg gcgccgcggg gcaccgcggc 63720 gggcacccgg ccgatgcgcctgcgcgacca ggccgacgac ctggcggcgg tgctcgccac 63780 cgaaggcgcc cgggactgccatctggtgtg ctggtgcagc gggcccaaga cggccatgga 63840 gttccgtcgg cgccgccccgagtccgtgcg gtcgatggtc tttctgcacg gctccttccg 63900 gcacacgggc gatccggccggcctcgaacc cgacacgccg tacgagcgca atctcgagga 63960 gctgtgccgc gccgtcgtcgcccggcccgc cctcgcggcc cggctgcgca ccatgttcac 64020 cggtcaaccg gccgagatgtcaggggactt gaccgcccat cggttcgccg acgaggtcct 64080 cgcgctgccc gcgcccgccctgggcggcga gctgctgcgc cccttctcgg acgacgccgt 64140 cctggtcgcc tatgcccgccagctcctgga ctactgggcg tacgacggtc tcgcgcacgc 64200 ggccgacgta caggtaccggtgctgtgtgt gagcggcgag ttcgaccgga tcgcctcgcc 64260 cgagcggctc gccctggccgcggaccgctt tccgggcgcc cgccacgagc gcctggccgg 64320 cgccacgcac cactccatgtacgaccgccc gcgcgaggtc gcgcggctgc tcacggactt 64380 cttcacggag gcggttcggccgcgctcaat tgtgagcggc gccgcacggt gacgctgcct 64440 agcgtcggac accaggggtcccaaggcctc ttcaccagtg cgacgaagga gacagagcga 64500 tgaccgacgc ggcgagccaggcccagggcg cgtttcttcc cgcccacagc gggcctctcg 64560 gttccggctc agacctgctgcactccggga acgcgggctt catcatccac cgggccggcc 64620 agctcaacca cgcgttccgcgcccaggggc gtcagttcgc cacggatctc gtcggccacc 64680 tgaacaaggt ggtggagggcgtcgccacga tcgtggtgca cgaggagatc ctcggcaccg 64740 ccgaccggct ccactggctgatccacatga agcagcccaa cgactacagc cgcttcctgg 64800 agatagccga ccacgaccgcagcttcaagg agatcaccga ggccgaccgg atcgcggccg 64860 cggagggcgc cggcaactgggagcgcatgt tcgtcgaggg ctccttccag gagcgggtgt 64920 acgtccccca gcacggcctggacgagcacg acgacgacca cgaccacgac caccacgacg 64980 agccgtcgga caccttcgtgccgcccgccc gccaccagac cgggctgccc gacagccgga 65040 tgcgcagctc cgtggacagcggcctgacga tcatccgtac ggcgcagacc gcgttccgct 65100 tccgcaccga ggcccgcgagttcgccttcg cctgggcctc cgaggtcaac cgggccctcg 65160 gcggcgagct gacggtctacctctacgagg agacgttcgg ccagcaggac cggatccact 65220 ggatgatcca cctcgacagcctcgacacct accgcaagct gaccgagctg agccgccacg 65280 acgccgacta ccaggcgctgttcgggcggc agttcgtgcc cgacttcaag ggcggtggcg 65340 gctgggagca gacgttcgtgtcccccacca tccaggacac cgtgctcacg ccgctgcacc 65400 ccggcgcgcc cgcctggttctgagcgggac ggacgcagcc gtgacggcca tcggcccgac 65460 ccaccgcggc gtgcggctcactgccgagcc gcacgtcctg cgcgccactc tcaccagccc 65520 ggacggcctc aacagcctgagcggtgctgc cctggacgcg ctgggggccg ccctggaccg 65580 ggccgaggcc gaccccgagtgccgtgtcct gctcctcgag ggctcgggcg gcaccttctg 65640 tacgggcctc gacttcgaggaggccgccgg cgatcctgcg ggcggcgcct cgcaggccgg 65700 ccggggcggg gccgagttcctggctctgat gcggcgcttc ggcgagacgc cgctggcggt 65760 cgtggcctgc gtcgacggccgcgcggcggg tggcggcgtg ggactcgcgg ccgccgcgga 65820 cctggtcatc gccaccgagcgcagcgagtt cagcctgccg gaagcgctgt ggggcctggt 65880 gccctgctgt gtgctgcccgttctcgtacg gcgtaccggc ttccagcccg cgtacgcgat 65940 ggccctgagc acccagccggtgtccgcgcg ccgggccgcg gacttccgcc tggtggacga 66000 ggtggtgccg gacccggatgccgcggtgcg ccgcctcctg gtccgtctca cccggctcga 66060 cccggcgacg atcggcgagctcaagcagta cttccgggcc atgtggttca ccaccgagga 66120 caccgacgcg ttcgccctgcgtgagttcac ccggctcatc gactcgcccg tcgcgcgacg 66180 ccgtatcacc gactacacgacaacgcggcg gctcccctgg gagaagccgc ggccctgacc 66240 ggggccgaag ccaggagggacgaggcgatg gtggcactgg ttttcccggg acagggatcg 66300 cagcgcaagg gcatgggggccgacctgttc gcccgcttcc ccgacctgac gcggcaggcg 66360 gacacggtcc tcggccactcggtggaggaa ctgtgccgtt cttcgggcga cggcaggctc 66420 gaccgcaccg agtacgcgcagcccgccctg ttcgtggtca gcgccctgtc gtatctggcc 66480 cgggatccgg ggctgccccagccgacgttg ctggccgggc acagcctggg cgagtacggc 66540 gctctgttcg ccgcgggctgtttcgacttc gccaccgggg tacgcctcgt ccgcgaacgg 66600 ggcgccctga tggggcgcgcccagggcggt ggaatgctgg cggtgctcgg cgtcgacggc 66660 gacgaggtgc aggcgctgctcgccgggacc ggggcgcgcc aggtcgatgt cgcgaactac 66720 aacacgccca cccagacggtgctgtccggc ccgctggacg aactgcgcat ggtctccgcg 66780 gccctgggac aacggccgggggtgcgatgt gtgcccgtgc gggtgagcgc ggcgttccac 66840 tcccgccata tgcgccccgccgcgcaggaa ttcgcgacgt tcctgaccgg gttctccttc 66900 gccgatccgc accgcacggtcatctccagc gtgacggcac gcccgtacgg ggcggggcag 66960 gtcgccgagc tgctgtcccgtcagatcgag agccccgtgc ggtggtcgga gaccatggcc 67020 tacctcaggg agcgcgggacgacggagctg gaggagatgg gcccgggcaa ggtcctgacg 67080 ggcctgtgga agcagggccgtgccgacggg gcgaaggccc gggccgtggc gcccgcaccg 67140 gtagcggtcg tggccggtgtccccgcggcg gccgcggccc gggcgcccgc ttccccggcg 67200 cctgtggcgg cgcgggcagcgactgcggcg cccgcacgcc cgtcggaccc cgctcccccg 67260 gctccccgca ccgccccctcccccgcctcg cccgtgcccc ccgcctccgt ttcgcgcggg 67320 cagcgggccg aggaactgggcagtgccgag ttccggcagg actacgggat ccgttacgcc 67380 tatctcgccg gggccatgttccgcgggatc gcctcggcgg agctggtcat ccgcatgggg 67440 cgggccggtc tcatggggttcttcggggcc ggcgggctcg ggctcgacaa ggtggagagc 67500 gccctggtgc ggatcaaggacgcgctcggt ccggacggcc ggtacggcat gaacctgctg 67560 cactcgatcg acgacccggcgtacgagcac gccgtcgtcg acctgtgcct caagcacggt 67620 gtccacgacg tggaggccgccggattcacc caactcaccc cggccgtcgt gcagttccga 67680 ttctccggtg cccaccgggatgccgcgggc cgcgccgtcg ccgtacgccg ggtgctcgcc 67740 aaggtgtcgc ggcccgaggtcgcggccgcc ttcatggcgc ccgcgcccgc ggcgatcctg 67800 cgccggctga ccgccgacggacggctgacg cctcaggagg ccgagatcgc cgccgagctg 67860 ccggtcggcc aggacatctgtgtcgaggcc gactccgggg gccacacgga cggcggcgcg 67920 gccctgacgc tgctgccctcgatgatccgc catcgcgacg cggcgatggc ccggcacggt 67980 tacggccggc ggatccggatcggcgcggcc ggcggcatcg gcgctcccga ggccgtcgcc 68040 gccgcgttcg tcctcggcgccgacttcgtg ctgaccggca gcgtcaacca gtgctcgccc 68100 gaggcgggta cgtccgacgccgtcaaggac atcctggccg gtctcgatgt gcaggacacc 68160 gcgtacgcgc ccgcgggcgacatgttcgag atcggggccc gggtccaagt ggtccgcaag 68220 ggaacgctgt tcgcggcgcgcggcaacaag ctgtaccagc tctaccgcag ccacgacagc 68280 tgggagtcga tcgacgcgggaacccggcgc tccgtcgagg agacctactt caagcggccg 68340 ttcgccgagg tgtgggaggagacccgcgcc taccacctcg ggcgggggcg cgacgccgag 68400 atcgagaagg cggaccggctgcccaagcac cggatggccc tggcgttccg ctggtacttc 68460 gcccgctcgg tgcgctggggcctggagggc gaaccgacgc agaaggtcaa ctaccagatc 68520 cagtgcggcc cggcgatcggcgccttcaac cacgtggtgc gcggcaccgg cctcgaggac 68580 tggcgtcacc gccatgtcgacctgatcgcc gaacacctga tgacgggcgc cgccgacgtc 68640 ctcgcccgcc gctgaccgtgcgccgcccgg cagcggcgga cggcgggcgg cacacgtatc 68700 gatccaccga accctggagccaccatgacc accctgacct tccactcgac caactccggt 68760 gtgatcgtgg agcgcaccgcccagctcaag caggcccatc gggccgaggg ccgccggatc 68820 gccctggagc aggccgcgtacctcaacgac aagttcgcgg ggcagaccac ggtcaccgtg 68880 cacgaggaga cgttcggcgtccgggaccgg ctgcactggc tggtgcacct gcccaagctc 68940 agcggccacc gcgagatctcccggctgctc gacggcgacg aggggccggc cgcggacgcc 69000 gactggcacg ggatgttcgtcgacggctcg ttctcggaga cggcgctgat cccgcagcac 69060 tggggcatgt acggcaccgacgaggcgctg cccgagggca ccgtcatcga cgccgccgcg 69120 cccgacctgc gcgtcccgccggcccagcgg cagacctcga tgagcccgga gcgcaccctc 69180 aactcgtccg gcgcgggactgatgatccac cgcgtcgccc agcccaagta cgccttccgc 69240 gccgaggcca ggctcttcgcccgccgcatc accgagtcga tcaacaccag gctgcccggc 69300 atcgcgtcgt cgttcctgtacgaggaggcc ttcggcccgg ccgaccgggt gcactggctc 69360 atccacatga agtccgaggacacctactac gacctgatcg acatgcacat gcgcatggac 69420 gacgcgaccc gcgcgatctatctggacgag atcatcgcgc cggagaaggg cggcggcacc 69480 tggaaccggc tgttcgtcgaggagagcatg ggcgacatcg ccttctcccc ggtgccggac 69540 gcccgctaga gaaacgctcgcttcgccgcc tcaccgcctc gccacccgcg ggcctgctca 69600 cttctgagca ggcccgcgggtgcgctgttc ctagggtgcg tcaggtgcag caagccggtg 69660 tcttttctgg gggagcggatatggagcctg gaaccagccc gtcggggcag caggatgcgt 69720 ccggcctcgc cgcacgggtgacggccgtga tggtgcgggt gctgtccgcg gcccggcccg 69780 acgcgcagcc cgtgaccgccgcatgcgaac tgcggggcct cggcctcgat tcgatgacgg 69840 cggcccggct gtggctcgcggtgcagggcg agtgtgcggc ggatgtgccg ctgggctggc 69900 tcaccgaggc gaccacggtcggcgagtacg cccagcgggt cgccgaccac gcgtcgcagg 69960 cggtgccggt ccagggggcgggtgccgtcg gagcacaggt cgccgcggac ccggacgccc 70020 tgcacgagcc gttcccgctgacgccgctgc aggaggcgta tctcatcggc aaggagcccg 70080 agctgcaggc ggacgccgtgggctgccacc tgtaccgcga gttcgacgtg ccggcgctcg 70140 acaccgagcg gctgcgcgccgcctggcagc gcctcgtcga gcaccacgac atcctgcgcg 70200 ccaccgtcac cgaggacggccggcagcaga taaccgcgca ggccccgcgc tgggacctcg 70260 ccgtgcacgg gagcgccacgcgcgccgagt tcaccgagac cgcgaccgcc gtgcgcgccc 70320 ggatgtccca ccacctcttcccggcgggcc actgtccccc cttcgcgatc gaggtcaccc 70380 tcggccccga cggcaccggccgcgtccact tcggcatcga cgcgatagtc accgacggtc 70440 agggcctcga cctgctcaccgcgcagtggg aggcgtgtta cgccgacccg tcccacctcc 70500 ttcccgcacc gacggccccgctgtcggtgc gggactgcgt tgtggccctg gacgcggcgc 70560 ggcggaccga ggcccaccggcgtgacctcg accactgggt gcgccggctg cgtgaactgc 70620 cgggcgcgcc gggcctgttcacggcggacg cgccggagcg cggcaccggc ctgtcgtgtg 70680 tgcgccgcag cagccgcaccgcgcggctga cggccgccga gtggcggtca ctgcgcgcga 70740 gggccgagga gctggcggtgtcgccgacct cgctggtcct gaccgtgttc accgaggcgc 70800 tcgcccggca cggcgcccacgagccgttct ccctggtcgt caccaccagc aggcgccctc 70860 aactgccgcc ggaggccgaccacttggtgg gtccgttcac gggcaccacg ttcgtcgagg 70920 cggtgccccc gcagcagcacacgttcgagg aggccgcccg tctcacccac gagggcctgt 70980 ggcaggcact ggagcactccgcggtctgcg gggtcagcgc gcagcgggcg ctgcgcggcg 71040 gcggtcccgg gccgctgcccgtggtgttca ccagcatgct cgacgcggcg ggccggccga 71100 gggcgcgcgg gttcgcggcggcgccggtgt acgcggtcag ccagacctcc ggggtgtggc 71160 tcgaccacca gatgtgggagcaggacggcg cgctgcacct gcgctgggac accgctgacg 71220 ggtgcttcgc gcccggtgtggtggaggccg cgttcgcgtc gctgtgcaac gggctgcgcg 71280 cgctcgccgt cgcgggacccgtcgtcaccc gccccctgaa cgacctgcag caggcgtact 71340 tcgtggcgcg tgccgccggggagccggggc cgtggcgggg ctgccaggtc gtcgtcccgt 71400 acgacaccga cgagcgcgtggacccggtgc gtctggagtc ggccgtggtg cggctcgtcg 71460 aggcgtacga cgtgctgcgctcggcggtca cgcaggacgg cgtcgtggag gtgcgcgccg 71520 gggcgccgcg ccggtggacggtgcccgtcg tcgcgggcgg ctgccccgat gaggtgcgcg 71580 atgccatggc ggcggcgaacttcccgctcg gccggtaccc gcagttcgag gtgcgggtgg 71640 tgcgcggcga cgacggtgacacggtcctca tgtcgatgga cctcacgctc accgacgccc 71700 gcggcatcca tctgaccggccgtgaactga tgcgcctgta cgccgatccc gcggcggagc 71760 cccggcccgc cgaggcggctcgggactcgg cgcgcgacgc ggacgagcag gcccgctcgc 71820 gggcccactg gcaggaccggctgcgggcgc tgccgcccgg tgtgccgctg cccgggccgc 71880 gtgacgcgga cgggcccgaccgccgggtcc gcctggccgg cgcgccgctg gcgctgcggc 71940 cgctcaccga ccgctgcgccgagcacggtc tgagcctgga cgcggtgctg ctcaccgcgt 72000 tcaccgatgt cctcgcccgcacgtacggaa cggacttcgc cgtgccggtg gtccgctggg 72060 accacggtct cgacccgcagcggcccggcg agttcacggc gctgagctgg ctgccgtgcg 72120 cgccgcgcga gctgtccttcaccgcgcggg cccggaccta ccaggagggt ctggagcggg 72180 acgccgatgt gtccggcagcggtcttcccg agctgcggcg ggccgtggcg cgctccggtg 72240 gcgccgggta ccccgtcgtctacacgtgcg ccctggacct caccgaccgg ccgctgccgg 72300 ggtcggtccg ggccgggcagtggctgagct gtaccccgga cgtgttcctg gactgcatca 72360 ccaccgtcga cgccggccagctgcagctgg cctgggacgc cgtggacggc cgtgccccgc 72420 aaggcggctg gagcgagctgcacgccgagt accgccggtc cgtggcccgt ctcgccgacg 72480 acgcggcggc ctggcaggagccggccgggg gcgacacgtc gggcgcggac gacggcgagg 72540 tgcgcggcgc cgagctgcacaagatcctgc acgaatggaa cgacaccacc cgcgcgttcc 72600 ccgacgaccg gctgatgcaccagctcttcg aggagcaggc ggcccagcag cctcgggcgg 72660 aggcgctgcg ctggcgcggcggcggcacga tgacatacca ggagctgaac cggcgggcca 72720 accgcatcgc cgcccggctggccgccgagg acgtgggccc ggagacggtc gtcgccgtca 72780 gtgtgccgcg cgggcccatgatggtcgcgg tggtcctcgg gatcctgaag gccggcggtg 72840 tctatctgcc gatggagccgcatctgcccg ccgagcgggc cgccgtcatc ctcgaggagg 72900 cgcacgccga ggtcgtcgtcaccaccgccg accgcgaggg ctggccggtg cccgacggat 72960 acgcccgcgt gtgcgcggacgccgccgtgg agggccccca ccccgccgac gcggacaact 73020 gcccgcggcc cgtgacgcagccgcacaaca cggcgtacat catcttcacc tcgggcagca 73080 ccggccgccc caagggtgtggcggtcgcgc accggcccgt cctgaacctc atcaactggt 73140 gccggcgcac cttcgggttcgggcccgggg acatgggcct gtgtgtcacc tcgctcggct 73200 tcgacctgtc cgtgttcgacgtgttcggcc tgctcggtac gggtgccgcg ctctacatcg 73260 cggacgcgga gcagcagcgcgatcccgctc tcctcctgga cgtgctgatc gaggagccgg 73320 tcaccttctg gaactcggcgcccaccacgc tcgcccaggt aggtccgctg ctcgacacag 73380 tgggcacggc gggtaccggtgatctgcgcc tggtgttcct cagcggcgac ttcacgccgc 73440 tgcccctccc cgacgaggtgcgcgccgtct tcccgcgcgc cgacatgatc agcctgggcg 73500 gtgccaccga ggcgacggtgtggtccaact ggttccgcat cggggcgatc gacccggcct 73560 ggcgcagcat cccctacggcaggcccatcg acaacagccg ctaccacgtg ctcgacgagg 73620 cgctgcgccc ctgtcccgtcggcgtcgagg gggatctgta catcggcggt gagtgtctgg 73680 ccctcggcta cgtcaaccagcccgagctga ccgccgaccg gttcatcccc gacccgttcc 73740 acgaggaccc tcaggagcgcctctacaaga ccggtgacag ggccctgtac tacccggacg 73800 gcaacctgag cttccaggggcgggccgacg gacaggtgaa ggtccgcggg ttccgggtgg 73860 aactggccga gatcgagcaccggctgcggg cgcacgacgg ggtcaaggac gccgtggtcc 73920 tggcccggga ggacggctgcggggaccgca ccctcgtcgc gtacctcgtg gccctgccgg 73980 gttcggcgcc ttccgggcgtgaactgcgcg ggttcgcggg gcagacgctg ccggagtaca 74040 tggtgcccaa cttcatcggtttcctggccg ggttcccggc caccgccaac gggaagctgg 74100 accgggccgc gctgccgtggccgctggcca aggcacacct gacgccgccg gaccgctctg 74160 ccgacgccga tcccgtcggttcggaggccg aagcggccgt ccctgtgaag gagtcggcgg 74220 ctgagcagcc tgccgtcgcgggcggggccg ggccctcggt ttccgtgccg tcgcgcgacg 74280 agctgtgcgc ggagatcgccgatctgttcg cccaggccct cggggtcgag tcggtggacg 74340 ccgacaccga cctgtgggaccagggcgcga cctccttcac catggtccgg gtctcgggct 74400 ccctgcagcg ctcgtacaagcagcggttcc ccgtctccgc actgctcgac aacccgagcg 74460 tcagcgcgat cgccggctgggtccatgcgc agctgggggg cggcgccgac gcggagagca 74520 cggccgccgc cgaggcggagaccgcgacgt cggtcgatgc cgagaccacg gcgacgaccg 74580 tcacgcagac cacggcggcctccgacgaaa ggcccgattc cggccccgga ccggtggact 74640 tcttcgccac cgaggagcgcgagcgcttca agcgacagca ctggaaccgg cgccccgacg 74700 agcccgggct tcccgaagtgccgctgggcg acgcccggtt cgaggacgag ctccacgcct 74760 ggcgggccag ccgccgggacttcctcgacc agcccgtgcc ccaccggtcc ttctcgcgtc 74820 tgctcggcct gctgcgcgagaccaccggcg ccgacggcac cggcgccctg tacccgtcgg 74880 ccggcgacac ctactccgtccaggtctatc tgcacctgac gccggacgcc gtcgagggcc 74940 tcgacgccgg gctctactactacgacccgt cccggcactc gctgcgcctg ctgcggtccg 75000 gtgtcctgcc ggatcgcggggcccacttct actacaaccg gccggtgttc gaccggtccc 75060 ggttcggcat ctacctgttcggccagcgcc acggcatcga gccgctgtac gccgaggaga 75120 gcctgcgcta cctcacgctggaatccgggt acatgagcca gttgctgatg ctggggcagg 75180 ccgcccacgg tgtcggcctgtgtccgatcg gcgccctgaa caccgagcag ttgagccagt 75240 ggctcggcct ggacgagggccatgtcttcc tgcaggcctt cctcggcggc gcggcggaac 75300 acccgcagcg cacggcgggcggcacggtgc cgttcttcac cgagccgacc gacagcgacg 75360 gcaacagcgg gagcggcgacagcagcaccg tcatcaccga tgccgtggcg ccggtcagcg 75420 ccgccgccga ggacgccgacgccgaacccc cggcgcacac cgccgagccg gccgcggtga 75480 tcggcatggc cggccggctccccggggccg gggacctgga cgccttctgg gacaacctcg 75540 tctcgggccg taccgcgatcggcccggcgc ccgcgtcccg gccggagacg gcgccctcgg 75600 gggcccgtgc caccggaggtttcctgccgc acatcgaccg gttcgacagc ctgctcttcc 75660 atgtgtcacc gcaggaagcgccggccctcg acccgcaggc acggctgatg ctcgagagcg 75720 tctggcagtg cctggacgacgccgggcaca cagccgactc gctgcgcagg tccgccgggc 75780 gcgtcggggt gttcatcggctccatgtggc acgactaccg gcagcagggc gccgaccggt 75840 ggaacggggg agacagcgcggaggtcgcgg cgaccgcctc cgacatcgcc aaccgggtct 75900 cgcacttctt cgacttccgcggccccagcc tggccgtcga cacctcctgc tcgtcgtcgt 75960 tcgccgcgct gcacctcgcggtggagagcc tgcgtcgcgg cgagtgcgga gctgccgtcg 76020 tcggcgccgt caacctgctggcccacccct accactgggg cctgctcgac ggcctcgaac 76080 tgctcgcggc cgacgccccgccggccgcgt acgccgccga aggcagcggc tggcacccgg 76140 gcgagggcgt cggggtactgctgctgcgcc cggccgacgc cgcccgccgc gccaaggaca 76200 ccgtgcacgg cctgatcgagggcacccgga tcggccacgc gggccgcgcg ccgcgctacg 76260 gcgcgccgca caccgcggccctcgccgact cgctcgcccg ggccctggcc gacgcgtcgg 76320 tgatccccga cgaggtcgactacgtggagt gcgccgcggc gggcgcgggc atcgccgacg 76380 ccgccgagct ggaggcactgggctccgtac tcgcccgctg cgccggagcg tcgcccgttc 76440 cggtcggcac gctgaagcccaacatcgggc acctggaggc cgcttcgggt ctctcgcagc 76500 tcatcaaggt gttgctgcagatacgccacg ggcgcatcgc gcccacgctg gtgtcgggcg 76560 agctcagccc gctcgtggactgggacggcc tcccggtcga gctggtcgac actccgcgcg 76620 cgctcacgcc gcgcgccgccgacggccggg ccaccgtcct ggtcaacgcg gtgggcgcga 76680 ccggttcgta cggccacgtggtggtgcgag caccgcacgc gcacggcacc gggcccgccg 76740 cgcaggacgg actcgcaggggccggggccg cgccgtcggc gagtggtccg cgcaccgtcg 76800 tcctgtcggc cgcctcgcccgaggggctca cggcggcggc cgggcggctg cgcgaccatc 76860 tggccggggc gggccgcgccctgtgcctcg acgacgtggc ctggacgctg cagaccgggc 76920 gcgcgtcgct cggtcaccgtctgacgctgt ccgccgacgg cctggacggc gtgcgggcgg 76980 gcctgaccgc gttcctcgacgggcgcgcct gtccgggcct cgcgaccgcc gccgccgatc 77040 cggcgctcgc cggagtacccgccggcgcgc aggacctggc gcgggccgcc ggggactggc 77100 tgcgcgggca cgcggtggacttcgcccggc tgtggtccgc gcccgcccgc cgcgtcccgc 77160 tccccgtcca ggacttcacggtcctcgcgc aggagcggca ctggctcgcc gcgcccgcgg 77220 cccggcggcc ggacggtgcggcgggttccg caccggccgc gccggagtcc gggcagtccg 77280 cgccgccggc gtccccgcaggttcaggacg accgcgcgga ccgcgcccag gagcatgtcg 77340 ccgcctgctt cgccgaggtgagcggcatcc ccgccgagca gctgcacccg cgggtcccgc 77400 tggagcacta cggcctgagctcccgcctgg tcgcccggtt caacgagcgg ctgcgccagg 77460 acgtccaagg ggtgtccagcacggtcctgt tcgagtatcc ggacctggcc ggggtcgccg 77520 cgcatctggc ggcccaccacgagggcccct ggtcggccgc cccggacacg cagccgtcgc 77580 cgcccgtccc gtccccggacccgctccccg tgccccgtac gcccgcggcg gccctgggcg 77640 aaagtgccgc ggccgacggtcccgagccca tcgccgtcat cggcatcgcg ggccgctacc 77700 cgggggccgg ggacctggagacgttctgga gcaatctggc cgaaggcgtc gacagcgtcg 77760 gtcccctgcc tgccgagcgcgcccgcgacg gctggcccac cgagcagatg tggggcggct 77820 tcctcgacgg tgtggaccggttcgacgcgc tgttcttcgg gatcgcgccg cgcgacgccc 77880 agctgatgga tccgcaggagcggcagttcc tccaggtggt ctgggagacc ctggaggacg 77940 cgggctgcac cagggcccgcatacgggagc aactgggctc cgacgtcggc gtgttcgtcg 78000 gcacgatgta caacgagtaccccttcttcg gcgtcgagcg ctcgctggcc ggggagagcg 78060 ccgacaccgg gtccgccgtcgcgggcatcg ccaaccgggt ctcgtacttc ctggatctgc 78120 acggcccgag cctcgccgtggacaccatgt gctcgtcctc gctgaccgcg ctgcacctgg 78180 cggtggagag cctgcgccgcggcgagtgcg ccgcggccgt ggccggcggg gtgaacctct 78240 cgctgcatcc gcacaagttccgccagcaga cccgcctgaa gatgtcctcc agcgaccacc 78300 gctgccgcag cttcggcgcgggcggcgacg gcttcgtgcc cgccgagggt gtcggggcgg 78360 tgctgctcaa gccgctgtccgccgccgagg ccgacggcga ccgcatccac gccgtcatcc 78420 ggggcacggc cgtcaaccacggcggcaaga ccaacggtta catggtgccc aaccccgtgg 78480 cccagggcga tctggtccgtgcggcgctgc gccgcgcggg cgccgacccg gccaccatcg 78540 gctacgtgga ggcgcacggcaccggcaccc agctcggtga ccccgtggag atcaacggcc 78600 tgaaccgggc cttcgcgggcgcctcggtgg ccccggcgag ccgtgccatc ggctcggtga 78660 aggccaacat cgggcacgcggaggcggcgg ccggcatcgc gggcctcacc aaggtggtgc 78720 tccagctccg ccaccgccacctcgtgccgt cgctgcacac cgaggagctc aacgacgcgg 78780 tggactgggc gagttcaccgttcgaggtgg tccgcgaagg ccgcccgtgg gccccgctca 78840 ccggagcgga cggcgccccgctgccgcgcc gcgcgggcct cagcgccttc ggcgccggag 78900 gagccaacgc gcatgtcgtggtggaggagt acgtccccgg cacggcaccg gaacccaccg 78960 aacccggcgt acccggcgtactcgaaccgc agttgatcgt cctgtccgcg cacgacctcg 79020 gccggctgcg ggcactggccggacggctgc gggaccggct cggacgcgac gaccggcccg 79080 cgccggccct cgccgacgtggcacacaccc tgcagagcgg ccgggagccc ctgcgcgagc 79140 gggtcgcgct cgtggcgtacgacgtggccg ggctgtgccg cgccctcgac ctgttcgcct 79200 cgggcgacac cggcgcctgggtgcacggcc gcacccccgg cggcgccctg cccgacggcc 79260 cgaaggcggt gctcgatgctgccgccgacc gggacgcgga gctgctgcgc ctgggccggc 79320 actggacggg gggcggcacggtcgactggc cgggtctgca cccggtgcgc cgccgcctgg 79380 tctcgctgcc ctcgtacccgttcgccgagg accggcactg gctgcccgaa ccgcggaccg 79440 cggctccggc ggcagcggcagcaactctga cggaaccgtc cgggaccacg ctgtacggcc 79500 gcacctggcg ggctctgccgccgctcgccg ccgcggccgc tcccgcaccg tcgggccgtg 79560 tcctctgtgt cttctccgcacccggcgagc ccgtggcgcg cgccctggcc gcactgctcg 79620 gccccgaccg ggtcacgctcgtccgggcgg gcgccgacgc cgggaacggt gtccccggga 79680 tcaccggcat cggcgatgaggccgaggctg ccgcgttcgc gcaggggctg cgggccgacg 79740 ggccggatgc cgtgggcggtctgatcgacc tcaccgacct cggcggcccc gcccacgggg 79800 acgccggatc ctggacggcccggctcgtcc tgctgcggcg gctcgtgcgc accctgcgcg 79860 gccacggcgg ccgcgtactgcacgtgaccg agggcctgta cgggcccgcg ggccccgcac 79920 cctcgctcgc cggtgcgcgcatggccggtt tcgtgcggat gctcggcgcg gagtacgggc 79980 gggtcaccgg caccgtgctcgatctggacg tctcggccgt cggacccgac gcggccgcgc 80040 gccagatcct cgccgagtacaccgggccgt acggcccggg cgacgtgtcc gtgcgcggcg 80100 gggtgcgcca ccgccccgaactcgtggccc tgcccgacgc cgggcaccgg tcgctgaccc 80160 ccgccgtgga ccgcgcctacctcgtcaccg gcggcacccg gggcatcggg gcccgggtgg 80220 cccggctcct ggtgcgccgcggcgcccggc ggatcgccct gaccggggcc cgtccgcagc 80280 cgccgcgtgc cgactggccgctcctttcgc cgggcacacc cgaggccgaa acggcgtcgc 80340 tggtagccga gttggaggcccagggcgcgc gcgtcctggt gcactccggc ccgctgagcg 80400 agcgggagcg caccgaccggttcctgcgcg aggtgcgcga ggtgctcggc ccgatcggcg 80460 gtgtcgtgca ctgcgcggggcgcgggcctg tcggcaggcc ctcgttcatc ggcaaggaac 80520 tcgccgactt cgaccccgtactcgaaccca agacgacggg cctcgaggtc ctcgacgagc 80580 tgtgcgccgg tgaccggccggagttcttcg tgctgttctc ctcgctgtcg gccgtcgcac 80640 ccggcctggc cgccggcgtcctcgactacg cggcggccaa cgccttcctc gactgttatg 80700 ccgatcacca ggtgcgctcgggacgcccct ggttccgttc ggtcgcctgg cccacctgga 80760 gcgagtcggg catgggcgcggaccggccgg acagctgcgc gcccgtgggc gtgggaccgc 80820 tcggcgacga ggaggggctgcgggtcctcg aacggatcct cgccctgccg gccgagcagg 80880 cccgcatcgt gccctgccccccgatcgacg gcatcgccgc cgaccccgcg gcactgctcg 80940 gttccccgcg ggacaccgacgcaacggcct ccgtggggtc cacgacgtcg gccgggtcca 81000 cacccatggc cgggtccacacccatggccg gttccacacc cgcagccggg tccgcacccg 81060 ttccgacgac gaccggggccacgccgcccc ggccccgtga agaggagcac accgtgccga 81120 acacctcagt caccgggccgccctggctcg ccccgctgtt cagcgagctc ctcgcgattc 81180 ccgaggacgc cctggacccgacggcgctcc tcggcgacct gggggtggaa tccgtcctcc 81240 tgggtgagat cctgctgcggctcgaggaac tgacaggtct gtccctcgac ccggcgacgc 81300 ttctcgacca ccccaccctcgaactcctgg gccggcatct ggcggacctc ggggtgccga 81360 gcgcaccgcc cgcgcccgccgcgaccaccg cgcccgccgt ggctcctgtg acccccgtga 81420 ctcctacggc tcccgtggctgtggcccctg tgacccccgt ggccccgtcc gggaagatcg 81480 ccgtcatcgg gctctcctgccgcttccccg gtgccgagga cgccgccgcc ttcgcacgca 81540 atctgctggg cggcacctgctcggtgaccg aggtgccgcc ctcgcgctgg gacgtcggtg 81600 agctgtaccg gcccgagctcgagccgggcc gcagcaccag caaatggggc ggattcctgg 81660 acgggatcga ggacttcgaccccgagtggt tcggcatgag cgaggacgag gcccgctgcc 81720 tcgacccggc cgtacggctgttcctggaag gctccgccac ctgtctgacg gacgccggat 81780 acggtgcgcg ggagctggcgggccgcgacg tgggggtgtt cgcgggcgcc cggatgtccc 81840 actacgggcg ccgggtgggcgagcggcgcg gtctcgtggg catgggctcc gaccagaact 81900 tcatcgccgc gcgcatcgcccaccacttcg acctgcacgg ccccaacctc gtggtggaca 81960 gcgcctgttc gtcctcactggtggcgctcc agctggcctg ccgcagtctc ctggacggcg 82020 agtcggagct ggcgctggccggcggggtcg acgtactcct ggacgaggag ccgtatctcg 82080 acttcagcgc ggccaaggcgctgtcccggc acggccgctg cgccaccttc gacgaggacg 82140 ccgacgggtt cgtgcccggtgagggctgcg gcgtcgtcct gctcaagccc ctggagaagg 82200 cgctgcggga cggcgaccggatccatgccg tgatcgacgc ggtcgccgtc aacaacgacg 82260 gccgcaccat gggactgaccacccccaacc cggcggcaca ggccaaggtc gtccgccggg 82320 ccctcgccgc ggccgggcggcgggccgacg aggtcggcct gatcgaggcc cacggcaccg 82380 gcacgatgat cggtgatccgatcgaactgc gtgccctgac cgaggtgttc cgcgaggaga 82440 ccggacggac cggcttctgcgccatcggaa gcgtcaagac caacgtcggg catctgctga 82500 gcgccgcggg catggccggcctgatcaagg cggtgctcgc ggtacgggac gggcggatcg 82560 cgccgacgct gttctgcgaacggcccaacc cgcgcttcga cttcgccgcc tcgccgttct 82620 acccgagccg cacggcccacgactgggtgc cggagcccgg ccgggtgcgg gtcgccgggg 82680 tgagcgcctt cggcctgggcggcaccaacg cccatgccgt cgtcagccag ttggatccag 82740 ttctggcggc ggcgcaccggccgcgcccgg ccctgcccgc gcccaacttc gcgcgccgca 82800 ggctgtggct ggaggcggcgcctccgtcgg cgcccggccc gcggccctcg gtccccggtc 82860 gcccgccgct gggtgcctccatcctcggcc tgaccttcga ggaacccctc tccgaacccg 82920 gttacgcccc tctcaccccaaccccgaccg gcgccgacac cgcagcggcg tcgcaggaag 82980 gacggaagtg gtgaacgtgccctccgcacc gcagcccgga cacgagttgg tgtgccatct 83040 cgtcctggag cacagcgacttcatcatgca gaaccaccgg gtgcacgggg tgtccgtcat 83100 gccgggggtg accttcctcgacatcgtctt ccgcatcctg cgtgaccggg gcttcgacac 83160 ggcccgtgcc gagctgcgcaacgtcctctt ccacgaggcg atcgccacca gcgagggctg 83220 tgaccgcgac atccgcatcacggtgtcgac gtccaccgac ggcagccgct ggatcaccgc 83280 ggagagccgc cgccgcgagggcggcgagag cgccgcggac taccaggaga acttccgcgg 83340 cgagctcgtc ctgcacgacgtgcccgagcc cgggccgctc gacgtcgacc ggctgcgcac 83400 caccgcgcgg cgcgtggccgacctggacga gatgtacgcg cgggcccgcg ccgaggagat 83460 ccggcacgga tccgcgatgcgctgcttcgg ccggctctac tacggcgacg gcgaactcct 83520 cgccgaactg ggtctggacggggaggcggc ggcgctcgac gagcacttcc atctgcaccc 83580 ggcgaaaatg gactgcgccaccatcgccgc gttcgcgcag gtgccgccgc cggaccagga 83640 cccgttcatc ccggtgttcatcgagtcgtt ccgggccccg cgcccgctga ccggcgtcgc 83700 gtacgcccat atgccgcgccccgagacgta cgcccagtcc ggggacatca tgcacaacga 83760 ctgcgcgctg tacgacgccgacggccgctt cctcgccggg ttcaccaagc tgacctgcaa 83820 gcgcatccgc aaccccgagctcatcacccg gctcctcgac gccccggacg tgacgcgcac 83880 ggccgcgccc gccccggctgcggtgtcccc ctcccccgtt gtcgcgcctg cctcgtccga 83940 cggcggggcc gggccggacgcggtgcgcgc gcatctgcgc gaactggtcg gcacccttct 84000 cggcagggcg ccccacgcgatccgcaccga tgccggtttc tacgacctcg gcctggactc 84060 cgggcacatg ctggacatcagccgccgcct ggaggaatac gtctgcgcgc cgctctatcc 84120 gacgttgctg ttcgagttctccgacatcga ctcgctcgcc gcgcacctgt acgccgagtt 84180 cggcgcgcag gtgcgcagcgcccccgcgaa cccgcctgcc acacctgcca cacccggtga 84240 ggacgccgga gcccctcccgcgtcggcggc caggagtacg gcccgcgccg tcgcgcccgc 84300 cctcggctgc caccggccggtgtggacacc gctgcccgcc gaccccggcg ctttcgcggc 84360 ggacggcgca cggaccgtggtgctcgtcgg tgccgacgcc gcgaccgcgg ccgccctgcg 84420 cgacgccgcg gccccggcccgggtggtacg cgccgaacgg gcctccgcgt tccagcgcct 84480 ggccgccgac cactaccggctcgacccggc cgatccggac cagctcgcct cgctgaccgc 84540 cgcgttggca accgacggcatatccgcgac ggcgtacgtc cgctgtgccc gcacccggga 84600 cacggacggc gccgggagcgcgctgccgga tgcctacctc gagagctggg cgctggccgt 84660 ggccgtcacc ggtacccggccgacggggcc tgtcccggta ctgttcctgc acccgcgcga 84720 tcccgccgcg ccccgcccgcacgaggacgc gctgggcgcg ctcgcccgca ccgtcgccgc 84780 cgaggcgccg cagctgcgctgccgtgcggt cggccacgac gccaccgcga cggccggcga 84840 tctcgccgcc gtcatcgccgccgagagcac ggacctgtcc gcggagagcg aggtacggca 84900 cacgggaggc acccggctcacggtgcgcca cgagacgctg gccgtgccct ccgggaacgg 84960 cgcgggcgtg ctgcgcgaggacggcgtgta cctcgtcacc ggcgggggcg gctctctcgc 85020 cgcactgctg gtggaccgcctggtgacgcg tggcccggtg cggctcgtgc tgaccggccg 85080 cagcgcgccg gggcccgagctcacacagcg gatcgaaggc tggcggcgac gcggcgccga 85140 ggtcacccac gtgcgcggcgacgtcgccca caccgacgac gtgctggccg ccgtgacctg 85200 tgcgcgcgag acgtacggccggatcgacgg ggtcttccac tgcgcgggca gcgtcgacga 85260 cggaatgttc ttccgcaaggacccggagcg ctccgcggcc gtcctggccg ccaaggtggc 85320 cggcacccgg aatctcgacgaggccaccgc ggacgacggc cttgcgttct tcgcgctgtt 85380 ctcgtccgtt tcggcgtccgtggccaaccc gggccaggcc gactacgcgt acggcaacgc 85440 gttcatggag cacttcgccgagcagcgtgc cgcccgcgcg gaccggcccg gcgtctcggt 85500 cgcggtcggc tggccgctgtgggccgacgg cgggatgcgg gtctccgaag acgtgctgcg 85560 ccgctccgcc gacacgtccgggctgcacgc cctgcccgcc gacgcgggcc tggacgcgct 85620 gttcgggctg ctctcgggggccgcgccgcg ggccgtcgtc acgtacggcg accaggagcg 85680 catcgccgaa ctgctgcccgcgccccggcc ctcggccgca cagtccgggc gcaccggctc 85740 ccccgactcg cctgactccccggacggcga cgacatcgcg atcatcggcg tcgcgggccg 85800 ctaccccgag gccgaggacctcgaggcgtt ctggcgcaat ctcgccgagg gccgcgactg 85860 tgtcggcgag gtgcccgcggaccgctggga ccacgcggcg tactacgacc cggagcgggg 85920 caaggagggc cgcacatacgggcgtcgggg cgggttcctc gacggcgtcg accggttcga 85980 cgcggcctcc ttcggcatctcgcggcgcga ggccgagctg atggacccgc aggagcggct 86040 gttcctcacc gtcggccggcaggccgtcga gaacgccggc taccgcccgg aagaactggc 86100 ccgcacccgg gtcggggtcttcgccggtgt catgtggaac cactaccagt tgtgtacgga 86160 cggcagtgcc gagcccgtcgcgccgaccgc cctgcactgc tccgtcgcca accggctctc 86220 gtactgcctg gatctgtccgggcccagcat ggcggtggac accgcctgct cgtcctcgct 86280 gacgtcactg cacctggccgtggagagcat ccggcgcggt gagtgcgcac tggcggtcgc 86340 gggcggcgtc aacgtcgccgcgcacccgca gaagtatctg cagctcgctc agggccggtt 86400 cctgtcgtcg gacgggcgctgtcgcgcctt cggcgcggac ggcgacggct acgtgcccgg 86460 tgagggcgtg ggcgcggtgctgctcaagcc gctggccgac gcgctggccg acggggacca 86520 cgtccacgcg gtgatcaagggctcgttcct caaccactcg gggcgcacca gcggtttcac 86580 cgtgccgagc cccgcggcgcaggccacgct gatcgccgac gcgctggacc gctccggtgt 86640 cgccgcggac agcgtcggctacatcgaggc gcacggcacc ggcaccgcgc tcggcgaccc 86700 gatcgagatc gaggggctgaggcaggcgtt cgcggacgcg gggctcgccc cgggcagctg 86760 cgccatcggc tcggtgaagtccgggatcgg gcacctggag agcgcggccg gtatcgccgc 86820 cgtcaccaag gtgctgctccagatgcgcca ccgggagctg gtgccgtcgc tgcacagcga 86880 acagcccaac ccgcacatcgacttcgcggc cacccccttc gccgtgcagc ggacccgggc 86940 cccttgggtg ccgcgaccgggcagcacggt gctgcgcgcc ggcgtgagcg cgttcggcgc 87000 cggtggctcc aacgcccatgtcctcctgga gagcgcgccg cccgcgccgg ccacccctgt 87060 ggcgggtccg cagctgttcgtgttctccgc caaggacgag cggacgctgc gcgaggtggt 87120 acgccgccag ctgcgccacctggacggtcc gggcccggtc gggagctcgg cggacgaggc 87180 gacggcgctg ctcaccggggaggtggccgc gctcctggac gtgcccgtgg acgccgtgga 87240 cgtgcgggag aacctggccgacctcggggt cgaccgcctg gcgctcgccg agctcggccg 87300 ccgcgtcgag gggcgcctgcccgccggggt gccgctgtcc gggcaggcgt cggtcaccga 87360 actggccgcg tccgcggcgctcgccgcccg gccggacgcg ctgccgctcg cggacatcgc 87420 gcacacgctg cgggtcgggcgttccccgct cgccgtcagg ctcgccgtcg tctgcggtga 87480 accggaggag ctgcgccggaggctggcggc gttcctggac ggcgacgagc ccggcgaggg 87540 cgtgttcacg ggccgcgccgacgacgacaa ggagcccgtg cggcttgagc gggccgccga 87600 gctgttccgc ctcggccggctgtccgagct cgcccgtgcc tgggccgacg gggccgcggt 87660 ggagtgggac gactgccgcgccggcgacgg cgtacggcca cgtcgcgtac cgctgcccgc 87720 gcaccccctg gacgagcgctcgtactggat cggcggctgg cgcagcgcgc aggagaccgg 87780 cgagagccgg gagccggtcgagacggcgca tgctccaact gccgtgcctg cggctccggt 87840 tgaccaggcg gacctgccggaggtgcggga ggagcggccc ggactcgacc cgcaggaggt 87900 gctgtgggcc gtggtggacgcggtccggac gcggctctat ctcgagcggg acgaggtcga 87960 ccaccggctg tccttcaacgagatgggcgt ggactcggtc ggcgccgtgg agatcgtcga 88020 gcagctcggc gcacgtttcgccctggagat ggacccggtg acgctcttcg accacccgac 88080 ggtgccccgg ctcgccgagcacgtcaggga actgcaccgg cagagtccgg cgccgcggcc 88140 acaggccgca ccggcccctgcacaaccggc ggcacccgag gccgccgccc tgcccgcacc 88200 cgcacccgca cccgcaccggcaccggcacc ggcaccggca tccaaacccg agccggccgc 88260 gtccccggac gcctgcgacgacatcgccgt catcggcatg tcgggacgtt tccccggggc 88320 cgaggacctg gacgccttctgggagaacat cgccgccggg cgtgacagct tcaccgaagt 88380 gcccgcgcag cgctgggacgtcggccccgt cttcgacgcc gaccgtctcg tccccgacag 88440 gacctacagc aagtgggcggccatgctgcc cgaggtgggc cggttcgacg cggcgttctt 88500 caaccactcg ccgctcgaggccgaggtgat ggacccccag cagcggctgt tcctcgagca 88560 gtcctgggcc gcactcgagcacgccgggta cgcggtgggc gccgacgacc gcacctcctg 88620 cggggtgttc gtcggctgtgcgcccggtga ctactcgaca ctgctcaccg aggcgggccg 88680 ggccgacacc ggccacgccttcctcggcac caccagttcg ctgctgcccg cccggatcgg 88740 ctacttcctg aacctggacggacccaccat ggccgtggac acggcctgtt cgtcctcgct 88800 cgtggccgtg cacctggccgccgacagcat ccggcgcggc gagtgcgcca tggccctcgc 88860 gggcggtgtg gccctcatggtcaccccgca gctgcacgtc agggccagca aggtcggaat 88920 gctgtcgccg cgcggcacctgtgtgccgtt cgacgcgtcg gcggacggca ccgtgctcgg 88980 cgagggcgtg ggcgccgtcgtactgaagcg cctggaccgg gccgtcgccg acggggacca 89040 catccacggc gtcatcaaggcgaccggggt gaacggcgac ggccgcacca acggcatcac 89100 cgcgccgagc gccctctcgcaggccgcgct catcgcggac gtgcaccgca gggccggggt 89160 gggcgccgac gacatcgggtacgtggaggc gcacggcacc ggtaccgcgc tcggcgaccc 89220 gatcgaagtg cgggccctgaccgaggtgtt ccgccgctcg accgatcgca gcggctactg 89280 cggcatcggc accgtgaaggccaacatcgg gcacaccacc atggccgcgg ggatcgcggg 89340 cctgctcaag acgctgctcgcactgcgcca cagcgagctg ccgcccgcgc ccgccttcga 89400 caccccgaac ccgaagacggagctggactc cagccccttc ttcgtggtcc gggaccgcca 89460 ggagtgggag ccgggtcccggcggtcagcg gatcgccacg gtcagcagct tcggcttcag 89520 cggcaccaac gcgcacgccgttctcgcgca ggcccccgag ccgcaggccc gccccgaaga 89580 gcccgaccag gagcggctgttcgcggtgtc cgcccgcgac ggcgcggccc tcgaccggct 89640 gctgctgcgg ctcgccgacagtgacctgga cggcgtcacc ccggccgacc tcgccttcac 89700 gctcggtgtc ggccgcgcccatctgccggt gcgggccgcc gtgatcgccc ggaacgtgcc 89760 cgaactgcgc cgcaggctccgcctgctgca gtccggcgcc caggccccgg gctgcttccg 89820 caccggtcag ggggccgcggccggcgacct ggacgagcag acccgcgccg agctcgcggg 89880 ccgcgcccgg tcgggttcgcccgccgagcg tgtcgccgcc ctggagcgcc tggccgccgc 89940 ctatgccgcg ggccaggacctcgactggca gtccctgtcg tacggggacc ggccgcgccg 90000 ggtgccgctg ccgacgtaccccttcggcgg ggaccggcac tggatcacac tccccgacac 90060 cgaccgcacc gccgtgcccgccacggcacc cgccacgtcc cgcgtcgacc tcccgggcca 90120 gtcaccgcag tcgactccccaccccctgct cggtgctgtc tccggggcgc ccggcgaccc 90180 ggacggcgcc cgcttccccgtgcccgtgcc ggccgcgcac tgggtgctcg accaccacag 90240 gatcggcgac cggccggtcctgcccggcgc cgccgggctc gacctcgcgg tggccgcggc 90300 ccggcgctgc ggcctgcgcggcaccgtgcg gctgcacggc gtgcagtggc tgcggctgat 90360 cgacgccgag gccgccggcacgctgcggct caccctgacc tcggacggcg aggggtaccg 90420 cttcgccctg tccaccggtgacgacgggac cgtctgctcc cgtggctcgc tcacggtgcg 90480 ccccgatgca caggacgccgccgccccggc cgcgtcgtcc gagaccctcg atgtcgcgga 90540 gatcgccgcc cgctgtccgtacgaggtccc cgccgagcgg ttctacgacg acttccgttc 90600 cggcgggatc gcctacggcccgtccttccg ggtcctggag aagatcacct ttggtgacga 90660 cgaggtgctc ggcacgctgcgcgccacccc ggacagcggc gggttcgccc tgcaccccgc 90720 cctgctcgac ggtgcccagcagaccatcgc cgccctggaa gggggcaacg acgccaccct 90780 ggtgccgttc tccgtcgagacggtggaggt cgtcgacgcg acggccgtcc ccgcgttcgc 90840 ccatgtcgtc cgtgcgggaaagcaccgcta cacggtgcgc ctggccgacc ggtccgggcg 90900 ggtctgtgtc cgctacgagggcctcgccct gcgggcgcag cacaacccgg tcgactccat 90960 gatgtaccgg ccggtgtggcgccccgctcc cctgccgcag cccggcaacg cgcccgcggg 91020 cggccgtacg gtggtggtccacaccgccga ctcgacgacc ctggccgccg cgctcgcggc 91080 ccgcacggga gccggcctggtggcgctgtc cggcgcgcag gacgcggcgc cggacccgta 91140 cgcggtactc gaacagcccctggaaaccgt ctacttcgtg gcccgcaccg gcgacgccga 91200 agggcccgcc gaggcggaccgcaccgccct tgacctgttc cgcctggtca agcggatgct 91260 cgcggtgggc cgtgcccgtgaccgcatcgc cctgcgcatc gtgcttgcgg gcgccgtacc 91320 ggcggacccc gaggacatgaccgagaccgt gcgtccgcac gccgccggag tgctcggcct 91380 ggcccgtgcg atcgagtcggagtgcccgcg ctggagcgtc gcctgtgtcg atgtcggcgc 91440 ggacggcggc accgtcggtgccgagcgcgc ggccgagcgg atcgtggccg agccgggcac 91500 cgaaccgctg gtgctgctgcgcggcgagga gcgcctggag cgcgtcttcg agccgctgcg 91560 ccccgccgcg ccccggggcacggagccctt ccgcgagggc ggcgtgtatg tgatcgtcgg 91620 cggcgccggc ggcatcggcttcgcgctcag ccggctgctc gcccggatcg cccgggcccg 91680 cctcgtgtgg atcggccgcagccccgaggg cccggaacac cgggccaagg ccgaggagat 91740 cgccgcgctg ggcggccaggtcctctacgt gcaggcggat gtcgccgacg aggcggcact 91800 gcgccggggg ctcgcctccgttcacacccg gttcggtcag gtcgacggcg ccgtgcacgc 91860 ggcgctcgac ctgcgggaccgcaccatcgc cctcatggac gaggaggact tcctcgccgg 91920 gctcgcgccc aaggtcgccggagtcaccgc cttcgcccgc gtcttcgggg ccgagccgct 91980 ggacttcatg ctggtcttctcctccgcggt gtccttcgtg gaggcgggcg gtcaggccaa 92040 ctacgcggcc gcgagcaccttcgaggacgc ctacgtccag tggctcgacc ggcggcacga 92100 ctacccggtg tccgtcgtgaactggggctt ctggggcagc gtcggcgcgg tcgccgacga 92160 ccggatgcgg gcggccttcgcccgtctcgg cgtcggctcc gtcgaacccg ccgagggcat 92220 ggcggtgctg cgccggatcatcgccgggcg gctgccgcag accctcgcga tgaaggcgga 92280 ccgcgcggcg ctgccggccatgggcatccg ggtcgcgggt acgcccgagg gcgaagggcc 92340 ggcccgtacg tcgccgcccgcgacgagcgc cgcgcccggc ccgggcacca cgaccccggc 92400 cccgtccccg gcccaggtcgcggtcccgga cccggtggct gccggtgcgg aggggctgtc 92460 ggccgaggcc gtgcggtcctacgtcgccgg ggtgttcgcg gaggtcctca agtacgagag 92520 cgcggcgctc gacccggaggccaccttcga gacgttcggc gtggactccc tggtgagcct 92580 caacatcgtc gaccgcttcgagcaggacct gggtgacctg ccgcagaccc tgctcttcga 92640 gtacatgacg atcgaccaggtcgccgggta cttccgccag gaacacggcg agcagctcac 92700 ccgggtcatc gccccggcccccacgcagga ccagggccag ggcgccgtgg ccgcgtcccc 92760 cgcggccgag gtggtgtccggcgccgccgt ccagggcggt gacgaggaag tgttcgcccg 92820 ggcgcaggag gcgttcgccgcggtcgaggc gttcagccgc gatctgctgc gccgcacctt 92880 cccgcgcttg gacggcgtgccccggcccgg cgagcggatc acggccgacg aactcgcctc 92940 ccggctcggc gtggtccgccggcaccgccg cctcttcgac gccgccctga gcatcctgcg 93000 gtcctgcgga gcggtgacgggcgacgcgga caccctcacg ttcgccgagc cgtcggcggc 93060 gccgggggcc cgcgtggagggcgccgaagt cgccgcgctc tacccggaga tgagcggtca 93120 cgtcactctt ctggaacgcactctcggcgc cctgggagag gtcctcgcgg gccgcaggaa 93180 cccgatggac gtgctgttcccgaagggctc cgtggccctg gtcgagccca tctacaaggg 93240 ccagcccatc gccgaccactacaaccggct gctcgccgac gaggtggccg acgccgcccg 93300 ccgcgtgcgc gcccaggaaggccgcccggt ccgggtcctg gagatcgggg cgggtaccgg 93360 agccagctcc cgtaccgtgctggccgccct ggccgccgcg gacgccgggg cccactactg 93420 ctacaccgac atctccccggccttcctgcg ccacggcgag cgggagttcg gccccaccta 93480 tccgcaactg gccttccacaccctcgacat ctcccgcgac ccggtggagc agggcatcga 93540 ggccgcctcg tacgacgtgatactcggcac ccatgtgctg cacgccacgc cggacatgga 93600 gcgcaccctg cgcaacatccgcaccctgct gcggccgggc ggactggtcc tcgtcaacga 93660 gatcacccgc ttctccgagttcctcacgct cacgttcggt ctgacgaccg gctggtggat 93720 gtacgaggac gcacagtgcaggctgccgca ctcaccgctg ctcgccccgg tccagtggcg 93780 gcagagcgcg gcggcggccgggctgcgcac cgtgcgcacc ggcggtctgc cgggcgtacc 93840 cgccgacgag ctggagcagtccctggtcgt tgcggagcgg cccgtcgagg actccggcga 93900 cgcgtcaccg gacggtgccgcggacgagca gtcgcccgag tccgtgcggt cctatgtgac 93960 cggcgtgttc gccgaggtgctcaagtaccg tgccgaggac ctggatccgg cggtgaccct 94020 cgagaacttc ggggtcgactcgctggtgag cctcaacata gtcgaccgcc tcgagcagga 94080 cctgggcgat ctgccgcagaccctgctctt cgagtacacg agcatcgaca gcatcgcgga 94140 gtacctgagc gccgagcacggggagcggct cgcccgggtc ctgggcgggg ctccggcggc 94200 ggcccaggcc cagccctcggctcccgtccc cgcccccgtt tccgtcgatg tccccgcccc 94260 catccccgct cccatccccgcccccgtttc cgtccccgtc gacgtccagg aacccgagtc 94320 cgagcccgag cccgcagcggctgtgcggac cccggccggc gacgaccccg cggatcctct 94380 cgacatcgcc gtcatcggcgtcgtgggccg ctacccccag tcccccgacc tggaggcgtt 94440 ctggcgcaat ctctccgagggccgcagctg catcaccgag atcccgtccg aacgctggga 94500 ctggcgccgc aacttcgacccggacaagag ccgcaagcac cgcagctaca gccgctgggg 94560 cggcttcctc gaggacatcgagatgttcga cgccccgctc ttcggcatcc tgccgcgcga 94620 cgcggccgac atcgacccgcaggagcggct gttcctcgag agctgctggg agctcctgga 94680 gacggccgga tacctcggcacgtacacgca cgaaccgcag accggtgtct tcgccgggct 94740 gatgtacggc gagtacggcctcctcgcggc ggccaccgac tggcccgagg gccgctacgc 94800 caccggccac tccgcgtactggtcgatggc caaccgtgtc tcttacacat tcgacctgca 94860 gggcccgagc ctcgccgtcgactccgcctg ctcgtccgcg ctcagcgcca tccagctggc 94920 ctgcgagagc ctgcgccgcggtgagagccg gatggcgatc gcgggcggta cgaacctgat 94980 cctgcacccg gcccacttcgcggccctgtg cgcccgcaac atgctctccg ccgccgacgc 95040 ctgccgcgtc ttcgacgacggcgccgacgg attcgtcccc ggcgagggcg cgggcgcggt 95100 gctcctcaag ccgctcgcgcaggccgaggc cgacggcgac accatctggg gggtcgtcaa 95160 gggcgcgttc agcaacgcgggcggcaaggt cagcggttac accgtgccca accccaacgc 95220 ccaggcccgg ctcgtggagcggacactgcg ccgctccggc gtgcacccgc ggacggtgtc 95280 gtacgtggag gcccacggcaccggcactgc gctcggcgac cccatcgaac tgggcggtct 95340 gaccaaggcg ttccgcgccgcgggcgccac cggcgacggc tactgcgccg tcggatcggt 95400 gaagtccaac atcggccacctggagggcgc ggccggtatc gccgccgtga cgaaggtgct 95460 gctccagctc aagcaccgcgctctggcgcc caccatccat ctggaccgtc tcaacccgaa 95520 gatcgatttc gcgggttcgccgttcgggcc gcagcgcacc gccgagccct gggaccgtcc 95580 ggtcgccggt gtggacggcgcggagcgcag ctggccgagg cgggccggca tcagctcgtt 95640 cggagcggga ggagcgaacgtacacatgat cctggaggag tacaccggcc aggacccgcg 95700 ggacgccgag gacaccatgggcgccgcggg tgccgaggag ccggagctgt tcgtgctctc 95760 ggcgctcgac cgcgaaaccctcgcccggca cgcggggcgt gtcgccgact tcgtggccgg 95820 tcccgaaggc gcgcgtgtccggctcgccga cctggcgcac acgagcaggg tggggcggcg 95880 cgagctgccc gagcgcctcgcggtgacggc cgcgtcgcat gcacaactcg ccgccagact 95940 gcgggagttc gccgcgaccggggtggcggg cgagggcgtg agcaccggca cggcccgcaa 96000 gggcggggcc ggcagcgggctcggtgccca ggaactcacc gccgccctgg ccgggcgccg 96060 ctgggcggac gccgtggagcactggacgct cggaggccgg gtcgactggc gtacggcgga 96120 cgcgggacgc ctggtccgcaaggtcgcgtt cccgacgtat ccgttcaacc gcagccgcca 96180 ctggatcacc tccgacccgcaccatctggc gccggagctt gcgagcggcg gccgtccggc 96240 cgaggagccg ctgtccggttcggagccggc ggcccccgaa ccggccgcga cccgcctccc 96300 ctccccggag ccggccggccccgaacccgt ggccgcagaa cgccccgtcg cggttcccct 96360 cccggtgccc gacctcgtgcagccggtcga ggacaacgat cgcgacgaag gcgggcccgc 96420 ggccgacgtc acggccgagcgcggggccac tcacccggcg ctcgccctcg cggcgcatca 96480 ccggatctcc ggtgcccgctgggtacccgg ggtgtcgctc ctggaagtgg cccgcgaggc 96540 ctgtcccggc gcggacggctccgcgggggc cgaattcccc ttccggctcg gctccgtgcg 96600 ctggctgacg cccgtcgacctggacgccgg gaccgaccgg gtgacggtcg gcttcaccga 96660 cgacggcacc gggaccgcgttccggatcac cgcgggcggc ccgtccggca ccaccgtcgc 96720 ccggggcacc ctcaccgagggcccctgcgc cgacgacgcc ggcatcgacc tggccgccgt 96780 gcgggaccgc agcaccgggcgccccgaccc ctgcgagttc tacgacgcgt tcgcccgcgc 96840 gggcctggag tacggccgccccctgcgcac cgtcgcggag ctgtggacgg gcaccggtga 96900 gtgtctggcc cggctgagcctgagccgcct gatgcgccgc gtcggccccg cccacgccct 96960 gcaccccgcg ctcctcgacggcgcgctgca ggcggtgtcg agcctgtgcg cggcggacga 97020 gtcgtatctg cccgtcggcatgggggagat acgggcccat ggcgcgctgc ccgcggacag 97080 ctgggcctac gtcagggaggtcacggccga gggcgggacg ggtggccggc gccggttcga 97140 catccggctc accgacggcaccgggcgggt cctgcaggcc gtggacggcc tggagatcgc 97200 cccgcgcgca ggaggcggcgcgcccgagac ccgtacccgc tacctggcac cggtgtggac 97260 gcccgcgcct ctgcccgccggggagcaggc gccgggcacc gtgctcctgt gcggcgccga 97320 gacgccccaa cgccaggaactcgccgggca gttgacggcc gcgggcagca ccgtggtgtg 97380 cgcgtccgac ggcccggccttcgcccgcag cgacgccggc ccccacgccg cgtacaccct 97440 gcgcggcacc gacccggccgagcacacccg gctgatcgag gacctcgcgg ggcgcgggct 97500 gctgccggat gccgtcgtacacctggccga cgaagcggaa gccgacatcc accgacagga 97560 cgtcgacagc ggtgagtcggtcctttcgtt cgtcctgtgg tccgcggtgg ccgtgctggg 97620 caggcccgaa cgccccggcctgcgcgtcgt gttcgcccac cgcgcgggtg cgcagggcgc 97680 cgagcccctg cacatcgcggtcggcgccac gctgcgcacc ctcggcctcg agcacagcag 97740 gttcagcggc gtacgcgtggagctgcccgc gggcggcgcc ggcaccgacg aggtcgtcgc 97800 cgagctgcgc gccgcgcggccgggtgccgt cgaggtctcg tacgccgccg acggtggccg 97860 cagcgagaag aacctggagccgttcgagcc gccggccgcc ggtctgtgcg acttcgcgcc 97920 gcgggcgggc ggcacctatctgatcaccgg cggcgcgggc gcgctcggtc tgcacttcgc 97980 ggcctacctc gccgggcgggtcccggacga cgccacgatc gtgctggccg gccgctcgca 98040 gccgggcggc gatgccgcccgccgcatcgc cgagctgaac ggcgacgggg cccgggtgct 98100 gcaccggccc gccgacctgcgggacgcggc gtccgtggag cgcctggtgg cgtccgtcag 98160 cgccgagttc gggccgctgcgcggtgtgct gcactccgcg ggcgtgaccc gggacgcccg 98220 ggcggtgaag aagacggcgcgggagctcga cgaggtcctc gcgccgaagg tgtccgggac 98280 cgtgcacctg gacgcggcgaccgcggacca gccgctggac ttcttcgtcc tgttctcctc 98340 gatgtccggc gagtcgggcaacctggggca ggccgactac gcgtacgcga acgcgttcct 98400 caacgccttc gccgcccgccgcgacggcat gcgcgcggcc gggctgcggt ccgggcgcag 98460 cctgtcgatc ggctggccgctgtgggccga cggcgggatg acggtggacg acgccacccg 98520 cgagatgttc gcccgcacctgggacatggt gccgctgagc acccacgcgg gtcttgaggc 98580 cttcgtccgg ggactcgcctgcgaactgcc cgccctgtgc gtggtggagt cgctggtacg 98640 gcgtgaggcg gcggccccggccacagcgcc gcggcccgct gccgggcggc aggagctgcc 98700 cgccatggac gtggtggcactgcgggagct cgtggagcac gaggcgcggg tgctggcctc 98760 cgggttcctg ctcgtggacccctcggaggt cgacgtggcg gccgaactgc tggagctggg 98820 cttcgactcg atcacactcaccgagctggt caacaaggtc aacgaacgct tcggcctcga 98880 cctgctgccc acggtcctgttcgagtgccc cgacctggtc tcgttcgcgg agtacctgac 98940 gcagcaccat gccgcggaactggcccagca ctacgcgccg gacagcccgg ccgagcgcac 99000 ggaagtacgc gacgagccggccgtgtccga cggcacaccc gccccggccc ccgcgtcacc 99060 ggtcgtcccc gcgtctcctgcgccatcggt cccgaccgtg accgcagtga ccacctcacc 99120 ggcacccgca ccggcaccggcaccggccgc ccgcgccgag atcgccgtca tcgggatcgc 99180 cggggtgttc cccggctcggcggacaccga cgagttctgg gagcacctcg cgggcggcgt 99240 cgacctggtg cggccggtgccgaaggaccg caccgcgatc cgcgccaacc cggccacccg 99300 cgagctgcgc ggtggcttcctcgactccgt cgacaccttc gatgcccgcc tcttcggcat 99360 ctcgcccaac gaggccgccctgatggaccc gcagcagcgg ctcttcctgc agaccgcgtg 99420 gcgggtgttc gaggacgcgggctaccgtcc ggccgacctc gcgggcgccc cgtgcggcct 99480 cttcgtcggg gtcgccacccatgactacga cgacctgctc aaggagaacg gcgtcgcggt 99540 acaggcgcac accgccaccggtatcgccca ctccgtcctc gccaaccggg tctcgtacct 99600 gttcgacctc aacggtcccagcgaggccgt cgacaccgcc tgttccagct ccctggtcgc 99660 gatccaccgc gccctgcgcgccatccagga cggcgagtgc gagctggccg tcgcgggtgg 99720 cgtcaacgtc atcctgaccccgggcctgct ggagtccttc acccagtcgg gcatgctcag 99780 cccggacggc cgctgcaagaccttcgacgc cgacgccgac ggctatgtcc gcggtgaggg 99840 cgtcggcgcg gtcctcctgaagccgctggc ccgggccgag gccgacggtg accacatcta 99900 cgcggtggtc aagggcaccgccgtcaacca cggcggccgc agcaactccc tgaccgcgcc 99960 caaccccgag tcccaggccagggtcgtggc cgccgccgtg cgcgaggccg gggtggaacc 100020 ggacaccatc acgtacatcgaggcgcacgg caccggtacc cggctcggcg acccgatcga 100080 gatcgagggt ctgaagaaggcgttcacgac gctgcacgag gagcgcggcg aggccgtgcc 100140 ggacaccggg cggatcgccatcggcgcggt gaagaccaac atcggtcacc tggagacggc 100200 ctccgggatc gcgggcgtcctcaaggtggt gcagagcatg cgccaccgcg tgctgcccgc 100260 gagcctgcat ctgcgccggctcagcccgta tctgcggctc gacggcacgc cgttcacggt 100320 caacgaccgc caccgcccgtgggagcccgc cctcacgccg gacggcaggc aggtgctgcg 100380 ggccggggtc agctcgttcggcttcggcgg ctccaacgcc catgtggtgc tggaggcgta 100440 cccggcgcgg accgcgcccgccgtacagga cttcgccccc cacaccgtgc cgctgtccgc 100500 cggcgacccg gacgatctgcggggttacgc ggcacggctc gcccggcacc tggcccgcac 100560 gccggaggcc gatctggcgcgggtcgccta cacgctgcag acgggccgga ccgggcaccg 100620 gcaccgcttc gccgtccgggtgcgggaccg tgacgagctg atcggcgccc tggaggcatt 100680 cgccgccggg gagctgccggaccacgcggc gaccggaacg gcccgccgtg acgctccctc 100740 cgtgcagagc gacgaggaccccgcgctact gcggaagtcg tggtgcgagg gcgccgacgt 100800 gccgtggcac acctggtggccgaagacccc gggacgtgtc ccgctgccca cggcgccgtt 100860 cgcccgcaca cgccactggttccccacccc ggatgccgaa ccgacggcgc ccgccgtacc 100920 cgcacgcaag gaggccaccatgacagcccg cagtgaggcg caaccgacgc gccgcggtcc 100980 caagatccgc ctcgcggcgccgaggagcgc ggggggtgcc tcggcgactg cgccgactcg 101040 ggaacccgtt cccgcgccgccggctccggc tccggctccc gccacgacgg caccgcagcc 101100 ggtccgggcc gacgtaccgcccccggccct ccccgaggcg gcggccgcct ccaccgtcgt 101160 cggcctgctg cgcggtgaactgtccaagat cctgggcatg ccgtccgagg agatcgagaa 101220 cgacgccccc ttcggcgagctgggcctcga ctccatctac cgcatggacc tcgtccgcac 101280 gctcaacgag gccttcggcctggacctgaa ggccaccgag ctctacaact acgacaccat 101340 cggcaagctc accgagttcgtcgcgccgct ggtcggcccc gccggggccg ctcccgccgc 101400 cgagccggtc atggccgaggcaccgcagca gtcctccgcg tcgctggagg acctggtaca 101460 ggacgtcatc gagcgtgagctcggccgcac cgccgacccg gccaagtcct tcgtcgacaa 101520 cggcttcggc tccttcgacatgctgcgggt cgtcgcctcc ctggagcggg tcttcggcgc 101580 cctgcgcaag acgctgctcttcgaccaccc cacgatcggc gcactggccg cgcatctcgc 101640 cgagacccac ggcccggaggccgccagtcg tctgtcgtcc ccgccgcagg accggccccg 101700 gcccgggccc gccgcgtcccaggagccgta caccggcggg gcgctagtgg tcgagaagaa 101760 ggccctcgcg ggtcagccggagctcgcctc ggcggtggcc ggtctggaga gcgcgtacgg 101820 ccgggagggc ggactgccgggccgcgacat cgccccgctg atcttcctgg gcgcgggccg 101880 cacggcgtac ttcaacttctcggtccggga cgacgccatg ctcacctgga gctacgtcgg 101940 cccgaccgag gagatgcccgccctggccac ggagttcgtg cgctacgggc aggcgcacgg 102000 cctggccgcg aacatcgtctcgctgatccg cctggaggag gtggacggcg tccggttcac 102060 cgccaccccc ttcggggcgctccaacgact cgaggggatc aaggacttca gcctcgaggg 102120 cggccggatg cagcggcttcgctacgcggt ccgcaagttc gagaaggccg gcacgtgccg 102180 gaccgaggag tacgccgtcggctccgaccc gcgtaccgac caggagatca ccacgctgat 102240 cgaccggtgg tcggcggccaaggagatggt gaacccgtac gtctcgactg tccgggacga 102300 gatcggccgc ggcattctcgccgcacggca ccggatgttc ctcacctatc tggacgaccg 102360 gatggtcagc gccgtcatcgtcaccaagat tccctccgag gacggctatc tgctcgacct 102420 cgagttctat cccgaggacgcgccgctcgg cagcctcgac cacaccgtcg tgaagatcat 102480 cgagcggacg gcggcggagggctgcaccgt cttcagcttc ggcggcagct tcggcgccaa 102540 ggtctgcgag tcgcccaacgccgcgccgga ggccgaggcc gccctcacgg agctgcgctc 102600 gcgcggcatc ttcaccggtgacggcaacct gcgtttcaag aacaagttcc gtacggagaa 102660 tctgccgctg tatctgtgccagcccgcgga cgcggagcgc accgatgtct cccggctgat 102720 cctgatgatc gccaacccggaggtgggcgg cgaccgcgcg cccacggccc cgaccgccct 102780 ggcagccccg gctccgcgcgaggcccctgc agcaccggct ccgcggcagg cccccgctcc 102840 cgggcaggcc ccggccgcccggccgaagcc cgccgccgct cccccggcgg ccgtcgcggc 102900 ccaggcaacc gaggtgcccgccgatgcccg tcgccgtgag cggcagctgg cggaccacgg 102960 ctggaacgcg ctccatctggcctccggcga cgtcgagttc gacctgatca ccgactcctg 103020 ggccgaactc gaccgccccttcgtgcacgc ccggacggcg cggctgcacg ccggggccgc 103080 cgggcgtccc gtgggccagacgctcgaagg gctcgatctg ctgccgttct cgtgtgtcgt 103140 ggccaccacg tccgggcgggccgcggaagc ggcgctgtgc cgggcctggc cgcagcaggg 103200 cgtcgtggtg cacaactcgctgttcccgac ctggtacttc aaccacctcg accacgggtt 103260 cacgcccgtc gcggcccgtcgcgcggccgg cgcggacgac ggtgtcttcc gcggcgacct 103320 cgacctcggc cacctgaacgggctgctcac cgaacacgcg ggccggatcg ccttcctgtg 103380 cgtggaggtc agcaacaacgcgcagggcgg ggccgcgctc agcctgcaca acctcaccgg 103440 catccgggag accgccgaccggcacggcct gcagctcgtc ctggacgcca cccgtgtcct 103500 ggacaacgcc gcgctgatcgccgcgcacga gccgggacag acgggccgcg atccgctcga 103560 cgtggcgcgc gagctgctctccctcgccga cagcgtcacc atcagcctct ccaaggactt 103620 cggcgtggac accggcgggatcgtcgccac cgacgacccg accgtcgccc accaccttcg 103680 ggagcggatc gcgctgcgcggcccggaggc gggccgtgcc acccgcgcgc tcgcggccgc 103740 cgcgctcgac gacctcggctgggccgcgac ggcgaccggc gaacgggtac gccgcgtggc 103800 ggacctgcgg caggccctcgccgcggccgg cgccccggtc gccccgggca ccggcacgca 103860 ctgcgtgctc ctcgacacggcgcgcctgcc cgcgctgcgc ggccatgagc acccggtgcc 103920 cgccttcctc gcctggctgtatctgcacac cggcatccgt gccgccgccc acctggacga 103980 cggtcccggc acctcgtccctggtgcgcct cgcgctgccg gtcgggctcg ggcagcggga 104040 gaccgccgag ctcaccgcccggctcacggc gctgttcggc gccccgcagc agatcccgga 104100 gctgctgctc gccgcctccgacgggcccgc cgccctcgcg tcgtaccacc cggtggagca 104160 ggtgcccgac gacatccgggaggcgatggc cgagggccac accgccgaga acgacaactg 104220 ggcggtgctg cgcgagcaccacccgggtgt cgagcgggtg ctgctgcggc tgcccgcggg 104280 cgacggcggc ggcgacgtcgaggtgttcac ggcgggcgcg ggcccggcgc tgctcatgat 104340 gcttcccttc aacatcggcgccgggctgtt cggcccccag ttcgcggcgc tcagcgagcg 104400 ctaccgggtg atcgtggtgcaccacccggg ggtgggtgac accaccgcgt gcgaggagct 104460 cggttacgag ggcatcgcggatctgtgcct gcgggcactg cgcaggctcg gcgtccaggg 104520 tcccgtgcac gtggcgggcgcgtccttcgg cgggatcacc gcccagacgt tcgccctgcg 104580 ccacccggag tccaccgcgtcactgaccct gatcggcagc tcgtacaagc tcggcaaccg 104640 ggccggagag gtcaaccggctcgccctggt ggccaaggag gacttcgacc aggtccagtc 104700 actcagcggt tcgtcgcggctggaccgtga acgcgcccgc ttcgagcggc tgctgctgcg 104760 ctgcgaaagc atggacccgcagaccggcct gcgctacctc gacgtgttcg ccaccgcgcc 104820 cgacctgctc ggccggctcggcgacatcgc cgtgcccacg ctgatcgtgc agggccggca 104880 cgacacggtc attccgcagaagacggcgca cctgctgcac ggtgccattc ccgacgcccg 104940 ctaccacgag gtgcccgacgccgggcactt cccgagcctg tcgagctcgg aggagttcaa 105000 cgccgtcctg tccgccttcctcgaggagca cccggcatga ccatcacctc gtcgctcgac 105060 gtcaggcccg agatcaaacaggccgtcacg gtgcggcccg gcatgtgcgg ccccggctcg 105120 ctgttcgtcg gccagctcggcgactggacc tgggagaccg tcagcgcaca gtgcgacacc 105180 gatgtgttcg cggcccgtgacgcctcgggc aaccccacct atctggcgtt ctactacttc 105240 cgggtgcgcg gcggacgggagctgcacccc ggctccctca ccttcggtga ccgcctcacc 105300 gtgacctcgg gctgctacgaccagggcacc gagtcggtgc tcacgctgca ccgcatcgac 105360 agggcgggca gcgatgacgcccaacgcccc ctggatctgc atgagttcta cgagcggccg 105420 cgggacgggt ccctgtacgtggagaacttc aaccgctggg tgacccgctc cgcgccgggc 105480 agcaacgagg acctggtgaagtcctcgccg cccgggttcc gcaatgacgg gctgccgcag 105540 ctgccggccg cgtactcgccgcgtgccgtg taccgcgagg cacgcaccgc gcacaccttc 105600 cgcgccctgg acgagcccggtttccgcctg ctgcccgaca ccgtcgaggt cgagcacccg 105660 gtggacatcg tccgtgacgtcaatggcgtc ggactcctct acttcgcctc gtacttctcc 105720 atggtcgaca aggccgcgctcgccctgtgg cgccggctcg gccgctccga ccgggcgttc 105780 ctgcgccggg tcgtggtcgaccagcagatg tgctacctcg gcaacgccga cctggactcg 105840 gtcctgaccc tcggcgcccgcgtccgggtc agcaccgaga cgcccggcga ggaactcgtc 105900 gacgtcgtca tcagcgaccgcgacagcggc cgcgtcatcg ccgtcagcac gctgcacacc 105960 cagcacgacg ctcacgacccgaagggggaa gcatgaccca ggcaccactg gatgtgcggg 106020 cacagacggc ccggctcgtcgtcgaggtcg tcaccgagat cctgcccggt gtggaccctc 106080 aactcatcgg cggcaagcggcatttgaagg acctgggggc cgactccgtg gaccgggtcg 106140 agatcatcgc ggccctcctggaccgcaccc gggtggacgc ccccatgtcc gacttcagcg 106200 acctgcccga catcgacagcctcatcgact ttctgagcgg agccccgcga tgaccgcgac 106260 cggtgccgaa caggcctatggcatcgaggc cttgaacgtg tggtgcggtc tggcccgcct 106320 caccgccgcg gacctcttcgcgggccgcgg cctcgacccc gagcgcctcg acaacctcat 106380 gatgtccgag cgctccatcggcctgcccat cgaggacccc gtcaccaacg ccgtgaacgc 106440 cgcccgcccc ctcatcgaggccctgtcgcc cgaggagcgc gcccggatcg agctggtggt 106500 gacgtccacc gagtcgggggtcgactacag caagtccctc acgtcgtacg tccacaagta 106560 cctgggcctg aaccggcactgccgcctcat cgaggtcaag caggcctgtt tcggcgcgac 106620 cgcggccgtg cagaccgcggtcggctatct ggcctccggg atctcgcccg gggcgaaggc 106680 cctggtcatc gccacggatgtggcggtggt ggacgagaag gccgagtact ccgagcccgc 106740 ggcagggcac ggcgccgcggccatgctcct gagtgaccgg ccccgcgtcc tcgccatgga 106800 cctcggcgcg ttcggcaactacagctacga gaccctcgac tcggcccggc cgtccccgcg 106860 cttcgacatc gccgatgtcgaccgctcgct cttcgcgtat ctcgactgcc tgaagaacgc 106920 gtacgcggac tacgccgcacgcgtgaccga tgtggacttc acccgcgact tcgaccatct 106980 cgtcatgcac accccgttcgcgggcctggt gaaggccggg caccgcaaga tgatgcgcga 107040 gcagggcgtc accgggccgcggatcgacga ggatttcgcc cgccgcgtcg ccccgtccct 107100 gatctacccg ggctcggtcggcaacctgtg ctccgggtcg gtgtatctgg ccctggccag 107160 cctgctcgac tccggtgtggtgaccgcgcc gagccgggtg ggcctgttct cgtacggctc 107220 cggctgctcg tcggagttcttcagcggaat cgtcgacgag cagtccgccg cgaccgtcgc 107280 cgaacagggc atcggcaagcgtctcgaagc ccgggcccgg atcaccttcg acgagtacct 107340 ggcggtcctc gaacacaacctcgagtgcct ggtgccggtc gagaaccgca ccgtggaccc 107400 ggccgaatgg gagccgctgctcgaccgggt cggcgaccgc ccggagatcc tcaccttcac 107460 cggcgtcaag gactaccaccgccagtacgc gtggcgctga ccccggcgcg gccgtccggc 107520 cgcggcccgc gaccacagcagcgacaagga gacgatgtgt acggctctcg gacggccacc 107580 cggcccgccg gcgaggacggggctgcggcc ggcctgcgcc tgttctgctt cgcccacgcg 107640 ggcggcggaa gctccttcttccacccgtgg cggcgggccc tggggcccgg cgtcgacgtg 107700 cgccccgtcg tcctgcccggccgtgagcgc cgggcgcggg agacctcgca cacccggatg 107760 ggccccctgg tcgaaggcctcgtcaccgaa ctggcccctc aactcgacct gccgtacgtc 107820 ctgttcgggc acagcctcggctcgatcgtc gcctacgaga ccgctcgggc cctgctcgag 107880 cggggctcac gcccgcctctggcgctgctc gtgtccgggc ggcgcgggcc cttcgtgccg 107940 gaccacaggc gcccggtgcacaacctgccg gaggacgagt tcctcgccga ggtgtcccgg 108000 ctcggcggca ccccgtccgaggtgctgcgt cagcgggacc tgctgcggca cttcctgccg 108060 ccgctgcgcg ccgaccacgaggtcaacgag acctaccggc cggtgccgcc gccgggaccg 108120 gccctgacct gcccggtgttcgcgttcacc ggcgacgccg accctctcgc cgacccccac 108180 gcggtggccc gatggcgcgaggtgaccagc ggcgacttcc ggctgcgggt ctttcccggc 108240 gaccacttct acctgaagggggcgccggac gacctgatgt ccgcgctccg ggcggccatg 108300 ggctccgtac atcagggctcaagtgttctt cgaaggcagg ttcataaagt tgccacgtga 108360 ttgtcatgtg ccgactgactgaccgggagg ccagatgaac ctgctggatg tcaccgaacc 108420 caccggattc gatgcccggctgcgtgactt cctgcgcgac cagtcgcccc acaccacgac 108480 gacgatggtc agacggaaccagagtgccta cagctgcggc ggtcaggacc gcaacgtgta 108540 cttcctggag agcggccggctcaagacggt gatgttctcc cgctcgggca aggagtgcct 108600 cctgcgcatc cacacccccgggagcatctt cggtgagctg tgttcgctcg gcgggatccg 108660 cgaggagtcg gccatcgcgatgcgcgacag cgtcgtccac cgcatgtcgt acgagcactt 108720 cctgtcctcg atcaccgaggccgggatcgt cgacgagttc atcaaccacc tcacccggcg 108780 cctcgccgaa cagcagcagtccatcacgca tctggtgacc gtcgacagcg agcagcggct 108840 cggcgagatc ctcctcgacctcgcccgcaa gctcggacgg ggcgacgggc ggcggccgca 108900 catcgcggag cggatcacccacgaggagct ggccggcatg gtcggcacca cccgctcgcg 108960 cgtcgggcac ttcctcaagggcttccgcga ccggggtctg gtcgaggtca cccgcgagtc 109020 ccacctgatc atcgaccagagccggctcgc cgcctatctg gagagccgat gcgtctgatc 109080 ccgccgcaac gccccgacgaccctcacgag gtgacacccc catgtgggac cacaagttcg 109140 agactctgct cagggaccggctgcccggtc tcgacgccga cgccccgctc accgaggatc 109200 tgccgctcgc ggccttcgggctcgactcga tggcgacggt cggcctgatc gtggccatcg 109260 aggaggacta cggcacgcccctgcccgacg acgccgtgac gcccgccaac ttcatcacgc 109320 cgggcacggt ctgggagctggtcagctccc tcgacggagc cggccgatga gcggcgccaa 109380 gctgctgcac cactggttcctcgacggtct ggcccgcaat ccgcggggca ccgcgctccg 109440 catcaacgac cgcagctggacctatgccga ggcggaccgc accgcgcgct cctgggccgc 109500 ggccctgcac agcggggggcgcgctccgcg ccgggtcggg gtcctcgcgg cgaagagcga 109560 ggagagcctg ctcggctttctggccgcgct gtacgcgggc tgcacggccg tcccgctcaa 109620 cccggagtac cccgtcggccgcaaccgcga catcgccgct gccgccgggc tcgacgcgct 109680 gatcgtcgac ccctcgggcgccgcccagct cgacgaggtg gccgcggccg cgccgccgct 109740 tgtcacgctg gcaccgcggctcggcgagtg gagcggaccg cggccgggca cccccgtcct 109800 gatgcccgac ccggcaggggaccggccccc cgctcagggc cccgactcac tggcgtacat 109860 cctgttcacc tcgggctccaccggccgccc caagggcgtg ccgatccggc acggcaatgt 109920 gtccgccttc ctggccgcctccctgccgcg ctacgacttc ggacccgacg acgtcttcgg 109980 gcaggtctac gagctcaccttcgacctgtc catgttcgag gtgtggtgcg cctggtcgag 110040 cggagcgtgt ctgaccgtcctcaaccggct tcaggccctc aacccgggcc gctacatccg 110100 tgctcacggc atcaccgtgtggacctcgac gcccagcctg gtcgccgcgc tgcgcactcg 110160 cggcctgctc ggcgggaacagcctgccttc cgtgcgccac accgtcttct gcggcgagcc 110220 gctgcccgag gagagcgccgcgtactggag cgcggcggca cccggcacgt ccatcgacaa 110280 cctctacggc cccaccgagctgaccatcgc ctgcaccgcg taccactggg tcccctcccc 110340 cggttcgggc accgtgcccatcggagagcc gaacaccggc ctccggtacg tccttctcga 110400 cgacggtcag gtcggcgccgacaccggcga actgtgcgtc accgggccgc agatgttcga 110460 cggctacctc gaccccgcgaacgacacgga ccgcttcctg acccacgacg gtcagcgctg 110520 gtaccggacc ggggaccgggtgcgccgcac gcccgacggc actttggtgc acctcggccg 110580 cgacgacgga caggtcaagatccacggcta tcgggtggag ctgtccgaag tggaggaggc 110640 cgtgcgggcc tgcgcgccgggcaccgaggc agtggtcctc gcggtcccgg gagcggccgg 110700 acccgtcctt gccgccttcgtcatcggcga cgacgcggcc ggccggctga ccgggacggc 110760 ccaggacctg gcccaccggctgccgccgta catgctgccc gcccatctgt gggcgctggc 110820 ggacccgccc ctgaaccccaacggcaagac ggaccgcctg cggctgcggg aagaggcggc 110880 acgccgcctg ggcaggaacctcgccccggc gtcctgaacc ggctgacggc acacggcagt 110940 tgacggtgga gcaggcccggatcagatccg ggtcagaccc cgtcgactgc cgtttcgcgt 111000 ggccgggact cgtccgccgccggtgtgcgc tccgcccggc cggccagttc tcctcgcacg 111060 gaggggaccg cgtccagcacggcccgggtc cacgggtgcc gcggggcggc gaacacctcg 111120 gcgaccgggc cctgttcgcgtacggtgccc ccgtcgagga cgaggacccg gtcggccatc 111180 cgcgcgatca cgcccaggtcgtggctgatc agcagcagcc cgaggcccag acgggagcgg 111240 agcccgtcga gcagggcgaggatctcggcc tggatgcgga cgtcgagact cgaggtgacc 111300 tcgtcgcaca gcaggagccgcggctgagcg gcgagcgccc gcgcgatggc cacccgctgg 111360 cgctggccgc ccgacagtgtgcgcggcagc cggccggccc actgcgggtc gagccccacc 111420 aggcgcagca gccgctccgtctccgcggcc gcggcctccc cgccgagccc acgcagcaca 111480 cgcagcggcc tggcaatggccgcacccacc gtgcgccgcg ggttgagcga tccgttcgcg 111540 tcctgcgcga cgagctgtacgaggcgacgc tggtcgaggg tgcggccgcg cagcggcccg 111600 gggacagcgg aaccggccagttcgatcagg ccgccgtcgg ggctgtggtg ccccgtgacg 111660 cagcgcgcca gggtcgtcttgccggtgccg gacccgccga gcagcgccac gcactcgccc 111720 ggctcgatgc ggaaggacacgtcctgggcg gtcgtcaccc gtgctctgcc gcgcccgtgt 111780 ccggcgcgca gcccgctgacccgcagcacg ggcgcggtgt ccgcggcgcc gccggccggg 111840 gcggccgggc gggccgccgcctcgccgatg tccggtaccg cgtgcaccag ttcgcgggtg 111900 tacgccgcac ggggccgcccgagcacggcg tccaccggtc cctcctccac gaccctgccc 111960 gccttcagca ccgcgagccggtcggtcagg cgggcggccg cgggcaggtc gtgcgtgatg 112020 aacagcagcg tcaggttccgctcggcggcc aggcgggcca cctcgtcgag gaccaggcgc 112080 tgggtgagca cgtcgagccccgtcgtcggc tcgtccagga cgagggcggc gggtccggcc 112140 gcgagcaccc ggccgagggcaaggcgctgc tgctgtccgc cggacagttc gtgcgggaaa 112200 cgacgcagga agtcccggccgtcgggcagg cccacgctgc gcagggtgcg ggccgcgtcc 112260 tcgggggtgc ggcgcacggcgagttccgag accagtgcgc cgacccgcat cgtgggggtc 112320 agcgcggcgg ccgggtcctgcggcagatac gccacggtgt gccgccgcag ccggcgcagg 112380 tcgcggccgc gcagggcgagcgggtcctct ccggccacgc gcaccccgcc gcccgcggtg 112440 cgcagtccgg gccgcagggcgccgagcagg gcgagcgcga gggtggtctt gcccgagccc 112500 gactcgccca cgacgccgacgcgttcgccc gcccgcacgg tcaggttcag cccgtcgagc 112560 acgggccgtc cgttcgggtcggccacggtc aggtcggtga cggagatgac ctccggcgcg 112620 gcgtcggccc gaccctgaccgtgcgggcgg gcgggggctg ggacggccag gtcgctccgg 112680 gcgtcgaaga ctgcgctcaaggcacgaacc tcgcgatcgc ccggtccagg gcaaggttga 112740 cggagacggt gaggacggcgagccccagtg cgggggccgc cacgccccag ggcgccagcg 112800 tcacgccctc catgttctcgctgatcatgg cgccccagtt ggggctgtcg ggtccgaagc 112860 cgaggaagcc ggcggacgcggtgagggtga tggcgatggc gaggcgggtg ccggcgtccg 112920 cgaggacggg ccgcacgatgttgggcacga tctcccgggc catcaccgcc accggaccgt 112980 ccccgagtgc gacggcctggtcgacgtatc cggcgtgcac gacctgtgcc gtcgcggccc 113040 gggcgaccct ggagacgaagggcacgccgg tgacggcgac cgccgtgacg agggtgagcg 113100 tgcggtagcc ccagccgtgcatcagcacca ggaggaccag gaccggcggc agggcgagca 113160 gcagggtgtc gaggcgggagaccaccgcgt cgacgcggcc gcccgcatag ccggcgagta 113220 ctccgctcgc ggtgccgaggagggtggtga cggcggtggc gagcagcggg atcaggacca 113280 gcgggcggcc tccggccaggaaccgggacc acacgtcgcg gccgtcgccg tccgagccga 113340 gcagcaggtg gccggtgtcgtagggcgcgg cggtgatgcg ggccgggtcg tgcggggcga 113400 gccagggtcc gatcagggcgaacaggacga cggccgccac gggcagcagc aacagcagcg 113460 cggcccgtcg gcccggtcggggtgcggcgg ggccggtgtc gggagcctgg gtgccggccg 113520 gcgtcaaagc ctcggtcgcagtcggcgtcg gggccgtcat gtcgccgtcc tcatgcgcgg 113580 gtccaccgcc atggccagcaggtcggccag gaggaggccg ccgacgaccg ccgcggccat 113640 gaggagcccg atgccttccaggacggccgc gtcgtgatgc tgcacggcgg cgaccaactc 113700 tcttgacaga cctgggtagttgaacacttt ttccaccacg gccgtgccgc cgacgagggt 113760 cccggcgagc cagccgaagacctgggcgca cggcccggcc gcggtcggca gcaggtgccg 113820 cagcaccacg cgggccggggacagcccggc gaggcgggcc gcctccacgt gcggggccgc 113880 cgccgcgtcg acgacggcggcccggaccat gcggctggcc caggacgcgg agaacagcgt 113940 cagcgccgcc accgggagcaccagcgcctc ggggtgtgcg agcaggggtt cgccgaacgg 114000 gaccagggac acggccgggagcactccgag ccaggaggag aagaccaggg tcaggatgcc 114060 cgccgtgacg aactggggcaccgagaccag ggtgacggcg ccgccggaga gcagccggtc 114120 gagcaggccg cccgggcgcagcccggcggc gattccgacc gcgagcgcga ggacgaccgt 114180 gagcagggcg cacacaccggcgaggagcag ggtgtcggcc agcgggcccg agatgatctc 114240 ggcgacaggg cggcggtccacgaggctgtg cccgaggtcg ccggagaacg ccgcgccgat 114300 ccagtcgccg tagcgcaccagcacgggccg gtcgaggccg agttcggcgt gcatcctggc 114360 gacgtccgcg ggggtcgcggagccgccgag cgcggcggcg gccgcgtcgc ccggcagcac 114420 ctcggcggcg gcgaacagcacgaacgagcc gaccagcagc gtggccaggg cgccgccgag 114480 ccggcgtacg gccagggccgcgagccggac ggcggacgtc acgcggacct ggcccgggtg 114540 aacaccggga tggactggctcgaccacacc cccgagagcc cgggcgccga ggcgtccaac 114600 tgctcctgga agccccacaggaggtcgccg ccggaggagt acaggcctcg ctggacggcg 114660 gcgaaggcgg cttcgcgcttgctgtcgtcg accgtggcct gggcgctcgc caggcgcttg 114720 aagtgcggct cgccgagcccggtcacgttg aagaacgcgc ccttggtggt cgtggcggac 114780 aggtgcacgg cggcgggccggttggcgtag tagaaggtct ggaacggggt gcgggcgagg 114840 cccttcaggt cggcgtagtaggtgtcggcg ggcaccttct gcacgtcgag ctccacgccg 114900 gcctccttca gctgctggccgagcaggcgg gccgcggcca cgaaaccggg cacggtgtcc 114960 gccgcgtgca gcgtcagcgagtggaccttg gcgcgggcga acagccgcct ggcctcggcc 115020 acgtcgcgct tgcggtccccgcccgcgtag ccgggcaggc ccttgccgac gacgtcgtcg 115080 cccttggtgc cgaggccgaggagcacctgc cgtacgagtg cctcgcggtc cgcggcgagg 115140 cgcaccgcgc gccgcacatcggggttgtcg aacggggcga gctgctcgtt catggcgaag 115200 gacagcgcgt tggagttggcggtgccgccc cggcggaccc ggatcgcgct gttgccgcgc 115260 tcggccttgg cgcccgtggcggtgatgccg aaggcgtagt cgatctggcc gctcttgagg 115320 gcggcgaggc gggcggcggggtcggcgatg aggtcgagtt cgatccgccg gaagtacgag 115380 gtgccgcccc agtacttgtcgttgacggcg aggcggaggt tcttgccgtc gtgctcttcg 115440 agacggtacg ggccggagccgacgcccttg tcgaagtcct tgccgcgggt gccttcgggg 115500 aacaccggtg agaagaccgcgaggacggcc tccctgaagt cgccgcgggg ccgcttgagc 115560 cccaccttca gggtgtgggcgtcgaccgcc ttgagccggt ccaggtccag gtcgacgaag 115620 ttgttggcgc gggtggacggcggcgtggcg agggtgcgca gcgagtaggc gacatccgcg 115680 gcccgcaccg gtgagccgtcgtggaacacc gcgtgcttgt ggaggcggac gatccactca 115740 ctggcgtcct tgttgggctccaccgactcg gcgagcccca gctcgtagcg gctgcccttg 115800 aggtacatca gctggtcgtacaggtggtac agggcgatgt agctggcacc gaacccggcg 115860 tgccgcgggt cgaggctggtggtgccggtg gcgtccgacg ccccgacccg cagcagcttg 115920 cccgcggagc cgaccgaccgggtgtcgccg ctcttggagg tggcgcagcc ggacagggcc 115980 gtggcgcccg cggtggcagtcgccgcggcc gcgccccact gcaggaagcg gcgtcggccg 116040 acgccgttga cccgtgacatggcagaaagc tcctcaggct catgaaggtg tgcgggagac 116100 gtacgtgggg ggcgggttcaggacctgccg gtcaggccca ccagtcgcgt gtgccgccct 116160 cgtcggcgcc gcattggcgccagatcttgc cctcgcgcag gacgtagacg ccgaaggtga 116220 cgaccgtctc gccgcggacgatcatggtgc cgcgcatcat gacggtgtcg tcggtctgga 116280 tgaactcgtt catgtccacgtgctggaggt cgagctccca gtaccgcttg agcagctcgc 116340 ggatgtcctc gcgcccgcgcacggggccgg tgaaccggat ccactcggcc tcggggtggt 116400 acaggtcgac gagcccgtcgaggtcccgct cggccgtgca gcggatcatc tcggcgaaca 116460 gcgggtggat ggggatctccgtgtcggtgc tcatgacggg gctcctgtcg cgtcgagttc 116520 tgcggatgcg gtgggtgcgggtgtcacgcc tgcggccgat gccgtcagcc ggtcccggat 116580 gagacgtttg acgaccttgccgaggtcgtt gcggggcaac tcgtcgacca ccaggaccct 116640 ttcggggatc tcgtggggggcgaggctgtc cttcgcgtga gcggcgaggg tggccgccag 116700 cagcttcggg tcatgggtggcgccggccgc gggcacgatc gcggcggcca cgccggccgg 116760 cccggcggcg accacggccgcgtcggcgac cgcctcgtgc tcgagcagga ccgactcgac 116820 ccgcagggac gacacgagcccggccggtgt gggcacggcg tcggtctcgc ggtcgaagag 116880 atacagctcg ccctcgtcgtcgacgtggcc gaggtcgccg gtgtcgcacc atccgccctc 116940 cgggacaccg gtggcctccgggcccgcgtc gaggcggttg cggggaggcg cgccgtcggc 117000 gcgcagccag atccggccgacccgatgggc gggcagctcc gcgccgtgct cgtcggtgat 117060 cagtacatgg gtgccgggggcgggccggcc ggccgacatc gggcgcgccg ggtcgaaggt 117120 gccgatggtc acggcgggcttggcctggct ggccgagtag gcgccgatca cgcgggcccg 117180 gggcatcgcg gccaacagggcgcgggcgag cgggggatgg aggaacgccg aggcggtggc 117240 cacggtggtg acggacgtcaggtcgtagcg gccgagcgcg ccgtcgcggg tcatctgcac 117300 ggcgaggttc ggggtcatcatgacggtgct cgccgcatgg tgctcgatga gcgcgcacag 117360 ccggtcggcg tctcccggcggcgcgaggac gagcgtggtc ggggtggcgt tgagcaccat 117420 ggagacggtg gtggcgctcgcggtgatgcc caggttcacg gacgccacca tgggggtggg 117480 ctcgtcgtgc gccatcacggagggcggtcc gcccgccgtc gcgaggttct ggtgcgagac 117540 gacgacgccg cgggccagtccggtggtgcc ggaggagtac acgatctcgg cggcgtccgc 117600 ggggctgtgg acgaggtccaccggtgtctc gcccggggcg gacagctcgt cgagcgtccc 117660 ggtccaggcg acggccgaggtcgtgggcgg ggcggtccgc ccgtggacga tgccggccag 117720 ctcgcactgc agggcacgccgttcgagctc gtcggccggg agggcgagcg gcaggtgaag 117780 gacggtggcg ccggccttgagcgcgccgag gtaggcgacc gcgtagccgg cccagtccat 117840 gccaccgaag aagatgccgatacgggtgcc ccggcttgtg ccggccgcga gcagcccgcg 117900 ggccagtgcc tccgagcgccggtcccactc gccgtacagc aggtcggggc agccgtcgat 117960 gacgagtgcg gtgcggtccgggtggagtgc ggcgtgggag cggagctggt cggtgaggta 118020 ggagaacgtc cgttccgtcatggcgtgctc aacccactat ctggaacgcg acaccgtcgc 118080 ggcccccgaa ccgcggcgcggtggcggcga gcaggtggcc gaggatgaac cgggcgcgct 118140 cctgcggcgg gtcgtcgctcagggatccga ggtagcggtc gtgggccgcg agggagcgta 118200 cggcccggtc gcgggagtcgtcgtcgacgg ccacggcgtg cgtggcgtgc ggggagccgg 118260 cgatcgccac cttgccggcccgccacggct cgagtccctc gtcgagcagc tcggggaaga 118320 tccagcggtt ggcggcgtcggcgacggcgt ccagggcggc gagacccacg gcacggtggt 118380 cgggggtgtt ccacgcgccggacgcccagg tgtcgtggtg gttgaaggtg acgatgagtt 118440 cggggcggtg ccgccggacggcgcgcgcca ggtcgcggcg caggccgggc ccgtactcga 118500 tcgtcccgtc gttgtagtggtcgaggaagt cgacctcgtg cacgccgagt tccgcggcgg 118560 ccttgcgctg ctcggcctcgcgcacgggcc cgcactgcgc cggttccaga tcggagatgc 118620 ctgcctcgcc ccgggtgacgaggaggtacg ccacctcgcg cccctgggcg gtccactggg 118680 ccacggcggg cccggcaccgaactcgatgt cgtcggggtg ggcgacgatc gcgagcacgc 118740 ggctgaagtc gtccggcagttcgaggagtg tgtcggccat ggggtgtcct tcccggtgga 118800 cgaaggagat gaggtcaggcggcggcgtcg gtctgcttgg gaacccggat gaccagggcg 118860 gcgatgagga tcgcggccgcggccaggccc gcgctgcaca ggaacgcgac ggtgtagccg 118920 tgcgtgccgc cgaggccgccgacggcctgc gtgaccgtgg tcagcaccgc gaggcccagg 118980 gaaccgccga ccaggatcgcggtgttgaag agccctgaaa cggctccggc gtccgcgggg 119040 tcgacacccg tggtggccagcgtgcccgcc ggcatcatgg tcaggcctag gccgatcccc 119100 gagatgatcg tcgggccgaggacggtgccg acgaagctgc ccgacgacgc ggcgaggccc 119160 agccacccca tgccgagcccgagacagagc aggccggtca tcagcatccg ggtcagcccg 119220 acgcgggcga ccgtcttcatacagacctgg gcgaacacga gcagggtggc gatcaggggc 119280 aggtacgcga ggccggtctgcagcgcggag aacccgagga tctcctgcag gtacagggtg 119340 acgaagaaga agtaactcatctgcgtggcg ccgaagatcg ccatgatggc gagcgcaccg 119400 ctggtgttgc gcagccggaagacctgcagc gggacgaacg ggtgcccttg gcggcgttcg 119460 acggcgagga acacggcgaacagcacggcc gcgaccacca gcaggccgcc gcgcagcgga 119520 gaggtccacg agcgctgcgtgtcgaggaac gcgaagacgg tcatcagcag cgcggcggtg 119580 aacgtcaccg cgcccgccacgtcgatcgag ggccgctcat cggcacggct ctccttgatc 119640 gtggtggcgg cgagccccacgaccagcgcc ccgatcggga cggcgacgaa gaagctgaac 119700 tcccagccgg ggcccgacgtgagcagtccg ccgacgatca gaccgaacag cgcgccggcc 119760 gcctgagcgc tcccccacagggcgaacgcc cgctgccgcg aaggcccttc ctcgaacgag 119820 gtgatgatca gcgagaggacgacgggggcg gtcagcgcgc cgcccatgcc ctgcaccgct 119880 cgcgcgatga ccatgaccgtcgggtcggtg ctgaatccgc agacgacgga cgccagcgtg 119940 aacacggtga cgccgatgatcagtacccgg cgcctgccga acaggtcacc gatgcggccg 120000 ccgagcggca gcagtccgccgtacaggagg ccgtaggcgt tgacgaccca gggcaggccc 120060 gactcggtga agtcgagatcgcgctggatg aagggcagcg ccacgttgac gatggtctgg 120120 tcgacctgga tgagcagctcggccatcagc agcgtgacga ggatcagccg ggcgacgagt 120180 gcgcccgtgg cggggctttgctcggcggcc tgccggccgt cgtcggtcct gacggtcatg 120240 gctctcctga tcttcgtgccacacccgccg cttgggcagg ctcggtcgag gccacgttag 120300 ggacggggag cccgagccggatgctcaata cagcgcatcg ccggtggggg atcgaaagac 120360 ctgccgcacc ggcgcgcggcgcctgcgaca caccgcgggg gttcgccccg gcccccgcgg 120420 tgtccgaccc cgtcgtcaccactcgacgtg catcacgcgc ggagcacgaa tgatcatgtc 120480 cgccttccat tcgacgtcgccgctgagccg caggcccggc atgccctcga cgagggcctt 120540 gagcgcctcc tgcagttccacccgggcgag gtgcgcgccc aggcagtgat gggcgccgtg 120600 gccgaagacc atgtgccggttgtcggcgcg gtcgatgacg acctcgtcgg cgttgtcgaa 120660 gacgaggccg tcgcggttcaccgcgctcgt gtccacgatg accggctcgc cctcgcgcac 120720 gaggacaccg cccaccgtcatgtcctcgag cgcatacctc ggagagatgc cgttcgcggc 120780 gaacgggacg aagcgcagcagttcctcgac ggccgcggga atcgcctcgg ggtccgcgca 120840 cagtcgcgtc cacagctcgcgacgctgctg gagcacatgg acgaagctcg agatctcact 120900 cgccgtcgtc tcgaatccgccgatgaggat cgagatgcac aggccgatga gctccggctc 120960 ggcgagccgg tcatgggcgtcgcgcgcctc gatgagggcg ctcgtcaggt cgtcccgcgg 121020 ctccgcacgg cgctccttgaccaggtcctc gaagtacttc tgcagatcca tgaagctctg 121080 cacacactct tcgtcggtcagttcgcccgt ggacagcgcc gcgtcgcacc atccccgcag 121140 ccgggtcctg tcgtcctccggtacgccgag gagctcgcag atcacggtga cggggatggc 121200 cagcgcgtac gagccgaccagctcggcggg cgcaccgccg gccttcatgt cggcgacgag 121260 cccggcggcg agctcgccgatccaggcccg cttggactcg atgcgccggg gggtgaaggc 121320 cttggagacg agcgtgcgcaggcgggtgtg gtccgggggg tccatcgtca ggatgctctc 121380 gggcacggcc cgcggcgtcatgcgcggctc gtcgcgctgg gtggccaggt gacgggagaa 121440 ccgccggtcg gtcaggacgaaacgtgcgtc gtcgtaccgc gtgacgagcc acgcgggctc 121500 accgaacggc agctgtacgcggatcgggtc cccgtcctcg aagagcttcg cgtacgactg 121560 cgagagctcg agcccctcgcttccggtgaa ggggtacgcg accggtgccg gcttctcggt 121620 ttccacttcg gtgctcactgtcgactgcct cctcggaaat gcccgccgtg acgcggcgtc 121680 acgctaacgg cccgggccgcggcggcctgc tcaatccgga gcatcgggcg gaagtaccgg 121740 gcgggatact ccgcacatgactcagatgcg gattcaggcc accttcgtcg tcgacgtctg 121800 ggacggcacc gacgacgagccggtcgacgg cggcccggtc accggtcgcg tcgaactgac 121860 caagacctac acggaaggcgatgtgaaggg ctcggccacc ggccacatgg tgaccaccca 121920 aggccccggc ggcgcggcctacgtggcgca ggagcgggtc accggaatca tgggcggccg 121980 caccggaacg ttcgtcctggagcaccgggc gacgcaggtc ccgggcaccg acccggtgac 122040 gtgggcgggc atcgtgccgggatccggcac cggcgaactg gccggcgtca gcggcgaggg 122100 ctcactcggc cacggcacgctcgacctcca ggtcgagttc gccaccgacg gggctgccta 122160 gtccgcggcg cgaagggccgttcagccctt cccggccgcc gccaggatct tctcggaggt 122220 gtggaagaac cgccccgtggcctgtacccc gaaggtcttc tcgatggcgg ggaccgggtt 122280 gacggggcgg ccgttgaccagccgccaccg cacataggcc gtcgccccgc aggcgccgag 122340 gacttcccag tcgcccttggcggagcgcag cgggaactcg ccgtccgtca gcgggcccga 122400 gctgtagacc atggagaaccgctcgtccgg ggccggctgc cggcccacga acggggacgc 122460 tgccagctcc gtgcgcagttcgtccgtcgt gcgcaggatc gtggggacct cgaacccgaa 122520 ggcctcggcg agaccggcctccacgcgctc gacgatgcgg gcgcggtcgt tccccgcgcc 122580 ggtgaagaag gcgttcccgctggcgatata ggtgcgcgcg cccatgaatc ccagctgggt 122640 gagcacctca cgcaggcgggccatctcgac cttgcgcccg ggaacgttga cggcccgcag 122700 gaaggcgatg tacgccggaccggtctgccc ggccggagca gtcgcatcgg tcgcatcgct 122760 cggaacggtc ctcatgtgcgtgcgggttcc ttcctcgttg ccacgtcgcg cccggccgga 122820 gccgggcgcc atgttctcacgcgtttcaga acgtccgctc ggccaccggg aagcggttgt 122880 gcgccgcgtt gcagcggatcagatcgtcga cgaacacgtt gagcagaggc cgttcgtccg 122940 cgtgccagtg gaagtggccgccggggaggc ggcgggcgcg gaaggagcgg cgggtgtagt 123000 cgcgccagcc gcccaggtccgcaccccgca ccgtgaggtc gtcctgcccc gccagcgccg 123060 tgacgggggc gtgcacagtgttgtcgagtc cttcctcgta cgactccgcg agcgcgaagt 123120 cggctcgcag caccgggagttggagcctcg ccaggccggc ctcgccgaac gtgccgggat 123180 ggatacgggg gtggtggccggcgcgggcca ggaactcctc gtccggcagg tgcccgatgg 123240 gctcctcgcg cgggggcaggtgaggcgccg ggtacccgga gacgtacagc cgctcggcga 123300 ccagttcgtg gtcccactccagacgggccg cgatctcgta ggccagtacg gcgccgaagc 123360 catggccgta cagggcgaagggccggtcgc acaggcgggc cacggtgggc accagatcgt 123420 gcagcagcgg ctccaggcggcggcgcggct cctcccgcat ccggctcccg cgccccggca 123480 gttcgagggg ggtgacggcgatgtgcggct ggagacgctc gcaccacggc agatagacgc 123540 ccgctgaggc gccggcgtacggcagacaga acaggcgcac cgacgtgccg gtcatgcgga 123600 ggccgccggg gtgggagtgagggcggtcga tacgtcgagc acggtggttc ccctctcggt 123660 ccacaagccc ctttcgcagggctgatgctg ccgccggacc ctacggagga aatgcgccaa 123720 ctcccgtttt ctgtagtgggtttggtggtc ggctccgagt gcttgtgcac cgcgggtcga 123780 tgtccgttgc ccgcaccgagcgaccccaag ggcgcgtgtg gcagatctcg agcgcctctc 123840 gagttgccgt ccatagcgtcccctcgggga cgaccaggac ccgagagagg agtccggatg 123900 accggcacgc tcgtgcgcggcgccgcccga acacgtccgc gtctgatggc ggtccacgcc 123960 caccccgacg acgagtccagcaaaggcgcg gcgacgttgg cgagatatgc cgccgagggc 124020 gtcgaggtgc tggtggtgacctgcaccgga ggggaacggg ggtccatcct caatccgggg 124080 tacccgcgcc cgcgcgggcccgtggacatg tccgtgctgc gcgcccggga gatggagcgg 124140 gcccggcgga ttctcggtgtgcggcactcc tggctcggat atgtggactc ggggctgcca 124200 cagggggacg aaccgctccctgcgacctgc ttcgcgcgcg tcgccccaca ggcggccacg 124260 gcccgtctgg tgcggctgatccggcagttc cggccggatg tcgtcacgac gtacgacgag 124320 gacggcggtt atccgcacccggaccacgtc atggcgcacc gcatcacggt ggcggccttc 124380 acggcggccg gtgacggtgcggcgcatcct cgtgcggggc ggccgtggca accgctcaag 124440 ctgtactaca accacggctcgcatccggcg cgcagcgcgg cgctgcaccg ggcgctgcgg 124500 gagcggggca gcacggcaccgtacgcgccg ctgcccaacg agccggtgcg gcagccggac 124560 atcaccacgc gggtgccgtgcgccgagtac ttcccactgc gggaccgggc gctgcgcgcc 124620 cacacctcac aggtggaaccgcacggcgtc tggttcgccg tgccgacgcc ggtgcagcag 124680 gaggtgtggc ccaccgaggactacaccctc gcgcgatcgc tggtgcgcac ggagttgccg 124740 gagagcgatc tgttcgccggaatcgccacg gcggccaccg tcggacgccg cggcgtcggc 124800 ccgcgtcaga tcagggcgcgccgccccgaa gggctccggg tgaagatgcc ggcggacgcg 124860 gcgagggccg cgctgcgggccgcctcgtcg ggcaggagcg ggtcgggttc ggctcgcagc 124920 aggaccagtt ggtgatagagcggcgcggtg gcggcgacga gcagggggcg tgcggcggtg 124980 tccggcggaa cgtccccgcgctcgacggcc cgctcgacgg tgacctcgca gcgcgtgtag 125040 cggtcgtccc agaaccggcgcagggcgcgg gcggcctgtt cggagcggaa cgacgcggcg 125100 atcagggccg cggtgaccgacggctgctcg gacagggccg tctggacgtc gtggttcagt 125160 gccgccaggt cgccgtacagcgtgccggtg tccgcgggct cccagtcgtc gcccatgccg 125220 gcgttcagga cgtcggcgagcagcccgccg acgtcacccc agcgccggta cacggtggtg 125280 cggtggactc ccgcgcggtccgcgacggcg tccaccgtga gtgcgtcgta gccgtgttcg 125340 accagctcgc tacggaccgcgtcgagcacc tgggcgcggt tgcgggcggt gcggccgccg 125400 gggcgtcgga ccgggggcgccggggcggcg gtggtggcgg tggcggagcc ggatgcggca 125460 tgtggggcgg cagagtcggtatccggtatt ccggttgctc ggggaggcgg tggcatgaca 125520 gcatcttaat gcaactgttgtcgtactagt ggggtcgtcg atgatcttcc gaagtgttcg 125580 cccggccccg cggccctcgccggccgctcc gtccccgaat gcttcggagt cctcacatgc 125640 ctacgcagat cagcgtccgctccgtcacca agtccttcga cgaccggctc gtcctcgacg 125700 cggtgacgtg ctcgctccccgcgggtgagc gcgccgggat catcggggag aacggctccg 125760 ggaagagcac cctgctgcgcctgctggccg ggcgcgacca gcccgaccag ggcacggtcg 125820 tcgtccaggc cgacggcggcgtcggctacc tcgcccagga cgaacagctc ccttcgcgtg 125880 ccaccgttca ggatgtgatcgaccgggccc tgggcgacct gcgcaccatg gaggagcgca 125940 tgcgtcgcct ggagacctcgatggccgacg gcgacgagag cggcatggcc gaatacggag 126000 acctgatgac ggtcttcgagctgcgcggcg gctacgacgc ggacgcccgc gtggagcgcg 126060 ccctgcacgg tctcggcctggacctcctcg accgcgaccg caccgtggac ggcctctccg 126120 gcggagagca ggtgcgcctgcgcctcgcgg ccgtcctggc ggccggtccc gaggtgctgc 126180 tcctcgacga gccgaccaaccatctcgacg aggacgccct gagctggctg gaggaccacc 126240 tgcgcacccg gcgtggcaccacggtggcgg tctcccacga ccggatgttc ctcgagcggg 126300 tggcgacgag cctgctggaggtggacgcgg acagccgcgg catcgtgcgg tacggcaacg 126360 gctacgccgg ctatctcgccgagaaggcgg cggcccgccg gcgcgccgcc caggcacacg 126420 cgcagtggca ggccgacgtggcccaactgc gcgaggcggc ggcgacgacc gcccggcggg 126480 tggcaccggg gcgagcgatgaaggacggca acaagatggc ctacgaccgg gcggcgggcc 126540 gcgtgcagca gtccctggccagcagggtgc gccaggccga ggaacggctg cgccgcctcc 126600 tcgccgaccc ggtgccctcccctccggaac ccctgcactt cgcgcccgta ctgcgggcgg 126660 caggcgtgca gggccccgtcctcgacgcgg ccggcatcgg tgtcgccggc cggctcgacc 126720 gcacggacct cacgatcgcggcgggcgagc gaatgctgat caccgggccg aacggggcgg 126780 gcaagaccac cttgctgcgcgtcctggccg acgagctggc gcccgacacc ggttcggtca 126840 cccgccgggg ccggatcggctatctcccgc aggaaccggg tcccggccga cccggcgaga 126900 gtctgctcgc cgccttcgcgcgcggccgct ccgggcagac cgccgagcac cgcgaacgcc 126960 tgctgtccct cggcctgttcgacgccgagc agctcacggc ctcggtgcgg ggcctgtcca 127020 ccgggcagcg ccagcgtctcgccctggcac ggctcgtgac gcagccgtcg gacgtgctgc 127080 tgctcgacga gcccaccaatcatctttcgc ccgcactcgt cgaggagctc gaagcagccc 127140 tggccggcta cgacggcgctctcgtggtcg tcagccacga ccgccggctc cgacggcgct 127200 ggcagggtgc tcagctcaccctgtcgacga atgcaccaca ggccgtccgg gtctgagaca 127260 cggacgtccg cacgcaccaactcgcaagga gtcacccatg cttcagtcca gcgccgtgtt 127320 caagcaggtg tccgtcgcggaccctcagtt ccgtccggcc cctacgacgt caccaccggg 127380 ccggacgacg cgctgtggttcaccctgatc cactccggag gcatcgcccg cctcacaccc 127440 gacggcgcgc tcgaccgtcatgcgctcgac tccccggcgt gcggcccctc gatcatcacg 127500 ccgggcccgg acggggccctgtggttcacc cgttgccagg accatcgcat cgggcggatg 127560 acggtgacag ggcacgcgacctctttcgcc gtcgcgacgc cggacagcgg cccctacggc 127620 atcgccgcgg gcggtgacggcgctctgtgg ttcacccagt tgcacacgga ccggatcggc 127680 cgcatcaccg tggacggccgggtgaccgag ttcccgttgc ccttcagggg cgcttttccc 127740 tcggccatca ccgcggggcccgaccaggca ctgtggttca cgctgaaccg ggcggacgcc 127800 gtggggcgta tggatctcggcggagaggtc acggtccacc cgctgcccac cgagcacgcg 127860 gcaccggtcg gcatcaccgtcggcacggac ggcgcgctgt ggttcgtcga gatcggcgcc 127920 ggtcaggtcg gacgggtgggcaccgacggc gtagtccggg agttccctct cccggaccgc 127980 gcgtgccggc cgcacgcgatcaccgccgac cccgccaggg gcgtctgctg gttcaccgag 128040 tgggctgccg accgcatcggatcgatcacg cccgacggcc acatcgagga gtacgaactc 128100 ccggaccgcg gcgcggaaccgcacggcctc acagtggcgc cggacggtgc gctgtacgtc 128160 gcactggaga cagggcggatactgcgcatg gagctctagg accggcttgt ccggcgggtc 128220 gtgcccacgg ccagtccttcccgcgaacgc tcttcgcccc cggccgacgg cgcgggggcg 128280 aagctctgtt cacagggtcttttcatcagg gctcttcacg gtgtcaccgc ttcaccacgg 128340 acctcctccg gtgtggtgtcgcgcagttca ccgtcgagca ggacccagcg ggtgatcccg 128400 atcgactcca ggaacggcacgtcgtggctg gccacgatca gggctccctc gtaggacgac 128460 agggccgagg tcagctggcgcacgctcgcc atgtccaggt tgttcgtcgg ctcgtccagc 128520 atcaggagct ggggcgccggctcggcgagc atcagcgcgg ccagggtcgc gcggaagcgt 128580 tcgccgccgg acagggtcgcggcctgctgg tcggcgcgtg cccccttgaa caggaagcgg 128640 gcgaggcggg accggatgcggttgttggtc gcctgcggtg cgaaccgcgc cacgttctcg 128700 gcgacgctca gggcctcgtcgagtacgtcg agccgctggg gaaggaagcg cagcggcacg 128760 tgtgccacgg cctccccctccacgggttcg agctctccgg tgatggtgcg caggagcgtc 128820 gtcttgcccg ccccgttgcgccccacgagc gcgatccgct cgggcccgcg cagctcgaac 128880 tcgccctgca cccgggcgccatgggcgagg tgcagttcgc gcagggtcag tacggtgcgc 128940 cccggcggca ccgcggtgtacggcaggtcg acatggatct cgtcgtcgtc gcgtaccgcg 129000 tccaccgcct cgtcgagccggtccttggcc tcggcgagtt tctcctcgtg catgatgcga 129060 tgcttgccgg cggcgacctgggcctgtcgt ttgcgctcgc ccatgacgac cttgggctcg 129120 cgcttggtgt cccacatcttctgtccgtag cgcttgcgcc gcgccaactt gacttgcgcg 129180 tccgccagtt cgcgcttctgcttcttcagg tcggactcgg cgacccgcac catccgctgt 129240 gctgcctcct gttcggcggccagggcctcc tcgtacgcgg acagattgcc tccgtaccag 129300 gtgacctcac cgcctcgcaggtcggcgatc tggtcgacgc gttccaggag ttcacggtcg 129360 tggctgacca ggaccagcacgccggaccac gcctcgaccg tggcgtacag ccgccgccgg 129420 gcccggaggt cgaggttgttggtgggctcg tcaaggagca gtacgtcggg cctgctgagg 129480 agcagtgcgg cgaggcgcaggagcaccgac tcgccgccgg acacctcgcc gatggtgcgg 129540 tccaggccga tgtggccgaggccgagcccg gacagcatgg cgacggcgcg ctcctcgacg 129600 tcccagtcgt cgccgactgcggtgaagtgt tcctcgctcg cgtcgcccgc ctcgatggcg 129660 tgcagtgcgg cgcgggcctcggcgatgccg agggccacgt cgacacgcag ggacgtgtcg 129720 agcgtgacgt tctgcgggagatagccgacc tctccggtca cccggacggt gccggaggcc 129780 ggggtcagct ccccggcgatcagcttcaac agggttgatt tccctgaccc gttgaggccg 129840 atgagcccgg tgcggccgggcccgacggcg acctgcaggt cctcgaagac gggcgtgccg 129900 tccggccagg cataggtcaggccggcgcag gtgatggagg tggggccgga gaagttcggg 129960 tgggtggtca tggggcctcgcggttgctcg gtgcgaacgg tggtgggggt gcaacgcgtt 130020 cgagacaccg tgggcgcggcgacgacggga ggagatgccg gattccggac cgcggcctga 130080 tgggcggcgg cgcggggagggggacggctc gtgacgtcga ggtcgcacac aggagcgtcc 130140 gcagtgccgg gatgtacggcgtgagctgcg tgacgcgacg gtgactcggg acctcagacg 130200 agcaacgtcc ttctcctatcgacggcagca gaatcgctat acagcgtata gcgatgggtg 130260 ggggatgtca acgactcgctctcgacgatg cactccatgg ctctgcgctc catgcgctca 130320 gtagcggtag tggtccggcttgtacgggcc gtcgacctct acgccgatgt acgaggcctg 130380 ctcggggcgc agcgtcgtcagcttcacgcc gagggcgtcg aggtgcaggc gcgcgacctt 130440 ctcgtcgagg tgcttgggaagcacgtagac gtcggtcggg tactcgtcgg gcttggtgaa 130500 cagctcgatc tgcgccagcgtctggtccgc gaacgagttg gacatcacga acgacgggtg 130560 gcccgtcgcg ttgcccaggttcagcagacg gccctcggac aggacgatga ggaccttgcc 130620 atcggggaac ttccaggtgtggacctgcgg cttgacctcg tccttgacga tgcccgggat 130680 ctgcgcgaga ccggccatgtcgatctcgtt gtcgaagtgg ccgatgttgc ccacgatcgc 130740 ctggtgcttc atcttggccatgtccgaggc catgatgatg tccttgttac cggtcgtggt 130800 gacgaagatg tccgccttgtcgacgacctc gtcgagcgtc gccacctggt aaccgtccat 130860 cgccgcctgc agcgcacagatcggatcgat ctcggtgatg atcacccgcg caccctggcc 130920 acgcagcgac tcggcacaccccttgcccac gtccccgtag ccgcacacga cggcggtctt 130980 gccgccgatc aggacatcggtggcacggtt gatgccgtcg atgagggagt gacggcagcc 131040 gtacttgttg tcgaacttcgacttcgtcac ggcgtcgttc acattgatcg ccgggaacag 131100 cagggtgccg tcgcggtgcatctcgtagag acggtggaca ccggtcgtgg tctcctccgt 131160 cacaccgcgg atctccgaggacatggccgt ccagtcgagt ccgctgcggt ccaacagggc 131220 gcggaccacg gccagttcctcgttctcggc ctcgggcagt atccccgtct tgcggtactc 131280 gacacccttg tggacgaggagggtggcatc gccgccgtcg tccaggatca tgttcggccc 131340 ggtgtggccg ggccaggtcagcgcctgctc cgtgcaccac cagtactcct ccagggtctc 131400 gcccttccag gcgaagaccgggatgcccgc ggcggcgatc gcggccgccg cgtggtcctg 131460 ggtggagtag atgttgcaggagacccagcg gacgtcggcg ccgagggcga cgagggtctc 131520 gatgaggacg gccgtctggaccgtcatgtg cagggacccg gtgatcctgg cacccgccag 131580 cggctgcgcc gccgcgtactcctcgcggat cgacatcagg ccgggcatct cgtgctcggc 131640 gagggtgatc tccttgcggccgaaggcggc gagggacagg tcggcgacct tgaagtccgt 131700 gaagtcagcg ggcggctgcgagggcatgtg tgctccttga ggtacggatg ctctggatgt 131760 tctcgtggat ctcgagcgtcgcggtggacc ggttgagggt gatgtagtgc agcccgggcg 131820 ctccctcggc gagcagccgttcggccatgg aggtggcgta ctccacgccg atgcgatgtc 131880 cctcggccgg gtcaccggcggccgcttcga ggcggtctgc cagtgcggcg gggaagcggg 131940 cgttggacag ttcggcgaagcggcggatct gccgcacgtc ggtggccggc atgatctccg 132000 ggatgatcgg ggtgtcgcagccggccgccg cgacccggtc acgcagccgc aaatagtcct 132060 ccacgtcgaa gaacatctgggtgatggcgt agtcggcgcc ggcccggcac ttggcgacga 132120 agtggctgat gtcgctgtcccagtcggccg agcggggatg ccgctcgggg aaggccgcga 132180 cgccgacgct gaagccgcccaggttcctga cgagccggac cagttcatgt gcgtgcgtga 132240 acccctgggg gtgcggcgtccatggcgcgt tggggtcgcc gggcgggtcg ccgcgcagga 132300 cgaggacatc gcgcacgcccgcgtccgcgt actggccgat gatgtggcgc agttcggcca 132360 ccgagtgccc cacggcggtcagatgggcga ccgggcgcag ggtggtctgg gcggcgatgc 132420 gcttggtgac ctcgacggtacggtcccggg aggagccgcc cgcgccgtag gtgacggaga 132480 cgaacgtcgg cgagagggcttcgaggcggc ggatcgcctg ccacagcatg cggctgccgg 132540 cctcggtcct gggcgggaagaattcgaagg agaaggtcgg ggtgccgctc gccagaatgt 132600 cgcgcagcgt gctcaattgcggcctccggc gttgaagtag ctcgcttcgg ggtggtgcag 132660 gacgatcgcg tcggtggactgctcggggtg cagctggaac tcctcggaca ggacgacgcc 132720 gatgcgttca ggtgcgagaagcgtggcgat cttggcccgg tccgcgaggt cggggcaggc 132780 cgggtagccg agcgagtagcggcagccctg gtactcggtt cggaacatgc cgtcgagcag 132840 gtcggggtca ccgccgccgatacccagctc ggagcggacg cgtgcgtgcc agtactcggc 132900 gagcgcctcg gccagctggacggacaggcc gtgcagttcc aggtagtcgc ggtacgagtt 132960 cgactcgaag agcttggcggtcgcctcgcc gatccgggag ccgacggtga cgacctggag 133020 gccgacgaca tcggtctcgcccgactcctc cgggcggaag aagtccgcga gacacaggtg 133080 ccggccgcgg cgctggcgggggaaggagaa ccgggacagc tccgagcccg cctcgtcgag 133140 gaggatgagg tcgtcgcccttggacacgca ccggaagtag ccgtggacca cggccgcttc 133200 cagcaggttg tccgtctgcagccggtcgag ccagccgcgc aggcgcgggc ggccttccgt 133260 ctccacgagc tcctcgtacgtgggcccgtc accggtacgt gcctgcttga ggccccactg 133320 ccccttgaac agggcgccctcgtccagcca ggacgcgtac tccttgagct ggatgccctt 133380 gatcacgcgg gtgccccagaagggcggctc cgggaccggg ttgtcggtgg acgtgtcgga 133440 gcgtgcgggc ccctcttcgggccgctcctc gatctgtacg ctgcccgcct tcacccggcg 133500 ctgcttgagc tccggcagtgtcgcgcccgg cacgccccgc ttcacggcga tcagggcatc 133560 catcagacgc aggccctcgaacgcgtcgcg cgcgtaacgg acttcgccct catagatctc 133620 gtggaggtcc tgctcgacatacgccctcgt cagcgccgcg ccgcccagaa tgacggggaa 133680 gtcggcggcc atcttgcgctggttgagctc ctccaggttc tccttcatga tcaccgtcga 133740 cttgacgagg agtccggacatgccgatcac gtcggcccgg tgctcctcag cggcctccag 133800 gatcgcggag acgggctgcttgatgcccag attcaccacg ttgtagccgt tgttcgacag 133860 aatgatgtcg acgagattcttgccgatgtc gtgaacgtcg ccacgcacgg tggccaggac 133920 aatggtgccc ttgccctccgcatcggactt ctccatgtgc ggctccagat acgccaccgc 133980 gttcttcatc acctcggcggactgcagcac gaacggcagc tgcatctgcc ccgacccgaa 134040 gagctcaccg acgaccttcataccctccag gagggtgtcg ttgacgatgt ccagcgccgg 134100 gcgggactgc agcgcctcgtcgaggtcggc ctccaggccg ttcttctcac cgtcgatgat 134160 gcggcgcttg aggcgctcgtccagcggcag cgcggccagc tcctcggcct tgccggcctt 134220 gagagacttc gcggtggcaccctcgaacag cgccatcaac ttctgcagcg ggtcgtagcc 134280 ctcggcacga cggtcatggatcaggtcgag cgccgtctgc acctcctcct cgctgaaccg 134340 cgcgatcggc aggatcttcgacgcgtgcac gatcgccgag tccaagcccg ccttcacaca 134400 ctcgtccagg aacaccgagttgaggaggat acgggcggcc ggattgagac cgaaggagat 134460 gttggacagg ccgagggtggtctgcacgtc cgggtagcgc ttcttcagct cccggatcgc 134520 gccgatggtc gcgatgccgtcgccgcgcga ctcctcctga cccgtacaga tcgtgaaggt 134580 caggcagtcg atgaggatgtccgactcatg gatgccccag ttgccggtca ggtcgtcgat 134640 cagacgctcg gcgatggcgaccttgtgctc gacggtgcgg gcctggccct cctcgtcgat 134700 cgtcagcgcg atcagagcggcgccgtgctc ctgcgcgagc cgcgtgacct tcgcgaaacg 134760 cgactcgggg ccgtcaccgtcctcgtagtt cacggagttg atgaccgcgc ggccaccgag 134820 cttctccagc ccggcccgcagaacctcgac ctcggtggag tccaggacga tcggcagcgt 134880 ggaggccgtg gcgaaacggccggccagctc ctccatgtcg gcgacgccgt cacggcccac 134940 gtagtccacg cacagatccagcatgtgcgc gccctcgcgg atctggtcgc gggccatctc 135000 cacgcagtcg tcccaacgggcctccagcat cgcctcgcgg aacttcttgg agccgttcgc 135060 gttcgtccgc tccccgatcgccatgtacgc ggtgtcctga cggaacggga ccgtctggta 135120 cagcgaggcg gcacccggctccgggcgcgg ctcgcgaacg gcgggctgta cgcccctcat 135180 ccgctcgacg agccggcgcaggtgctccgg ggtggtgccg cagcagccgc ccaccaggga 135240 caggccgtag ctgttgacgaagttctcctg cgcgtccgcc agttcggacg cggtcagcgg 135300 gaagtaggcg ccgtccttggtcagcaccgg cagtcccgca ttcggcatgc acatcaacgg 135360 cgtcgtcgag tgctgcgcgagatggcgcag gtgctcgctc atctcggccg ggccggtcga 135420 gcagttgagg ccgatcaggtcgatgcccag gggctccagg gccgtcaggg ccgccccgat 135480 ctccgagccg agcagcatcgtgccggtcgt ctcgaaggcc agggagcaga tcaggggcag 135540 tccgacgccc agtgactccatggcccggcg ggcgccgagc agaccggcct tggtctgcag 135600 cagatcctgt gtcgtctcgacgatccacta gttcgacc 135638 2 289 PRT Streptomyces atroolivaceus 2 MetGlu Met Ser Asp Thr Ser Arg Ala Ile Ala Ala Ala Ile Ser Ile 1 5 10 15Ala Ala Ser Leu Gly Leu Pro Ala Asp Asp Ala Ile Val Leu His Asn 20 25 30Ser Asn Lys Leu Ala Leu Arg Leu Thr Pro Ser Asp Val Leu Ala Arg 35 40 45Val Ala Pro Val Gly Gln Glu Val Ala His Phe Glu Val Glu Leu Ala 50 55 60Gln Arg Leu Ala Glu Phe Gly Ser Pro Val Gly Leu Leu Glu Pro Arg 65 70 7580 Val Asp Pro Arg Val Tyr Thr Arg Asp Gly Phe Ala Val Thr Leu Trp 85 9095 Thr Tyr Tyr Glu Pro Val Thr Pro His Val Ser Pro Ala Asp Tyr Ala 100105 110 Glu Ala Leu Glu Arg Leu His Ala Gly Met Arg Lys Val Asp Val Pro115 120 125 Ser Pro Ser Phe Met Glu Arg Ile Thr Glu Ala Gln Glu Val ValAla 130 135 140 Ser Leu Asp Leu Ser Pro Gly Leu Ala Asp Ala Asp Arg ValLeu Leu 145 150 155 160 Ser Gly Arg Leu Gly Ser Leu Arg Arg Ala Ile GluGly Arg Gly Ala 165 170 175 Val Glu Gln Leu Leu His Gly Glu Pro His ProGly Asn Val Leu Ser 180 185 190 Thr Lys Lys Gly Pro Leu Phe Ile Asp LeuGlu Thr Cys Cys Arg Gly 195 200 205 Pro Val Glu Phe Asp Leu Ala His AlaPro Glu Glu Val Cys Lys His 210 215 220 Tyr Pro Asn Val Asp Gln Gly LeuLeu Asp Asp Cys Arg Gln Leu Val 225 230 235 240 Leu Ala Met Leu Ala AlaTrp Arg Trp Glu Leu Gly Asp Gln Phe Pro 245 250 255 Asn Gly Lys Arg PheGly Glu Glu Phe Leu Arg Leu Leu Arg Glu Gly 260 265 270 Pro Pro Trp ProThr Phe Asp Ala Val Thr Arg Arg Leu Gly Ser Gly 275 280 285 Gln 3 502PRT Streptomyces atroolivaceus 3 Met Ser Asp Phe Ser Arg Arg Arg Leu LeuLys Thr Ser Ala Val Ala 1 5 10 15 Gly Ala Gly Ala Val Val Val Pro GlyVal Ala Ala Ala Ala Thr Pro 20 25 30 Ser Ala Gly Val Val Thr Glu Ala SerVal Asn Glu Ala Tyr Glu Ala 35 40 45 Gln Cys Ser Pro Ala Lys Leu Thr GlyArg Ile Val Arg Pro His Asp 50 55 60 Pro Gly Tyr Thr Asp Ala Arg Leu GlyTrp Asp Gln Leu Phe Ser His 65 70 75 80 Tyr Pro Leu Val Ile Val Phe AlaGln Asn Thr Gln Asp Val Val Asn 85 90 95 Ala Leu Thr Trp Ala Arg Gln AsnAsp Val Ala Leu Arg Ile Arg Ser 100 105 110 Gly Arg His Ser Leu Glu GlyTrp Ser Asn Val Asp Ser Gly Leu Val 115 120 125 Ile Asp Ile Ser Glu LeuLys Ser Val His Leu Asp Ser Ala Ala Arg 130 135 140 Val Ala Thr Val GlyAla Gly Leu Asn Gln Leu Glu Ala Val Thr Thr 145 150 155 160 Leu Ala LysArg Asn Phe Ala Val Thr Thr Gly Thr Glu Gly Thr Val 165 170 175 Gly LeuAla Gly Ala Thr Leu Gly Gly Gly Phe Gly Phe Leu Thr Arg 180 185 190 TrpLeu Gly Met Ala Cys Asp Ser Leu Ile Gly Ala Glu Val Val Val 195 200 205Ala Glu Gly Gly Glu Cys Ala Lys Val Ile Lys Ala Asp Leu Thr His 210 215220 Asn Ser Asp Leu Leu Trp Ala Leu Arg Gly Ala Gly Asn Gly Asn Phe 225230 235 240 Gly Ile Val Thr Ser Leu Thr Tyr Lys Val Ala Pro Leu Lys SerVal 245 250 255 Thr Tyr Val Gln Ala Thr Trp Asp Gly Ile Gly Asp Leu ArgArg Val 260 265 270 Phe Asn Ala Trp Gln Arg Thr Ala Leu His Ala Asp AspArg Leu Gly 275 280 285 Thr Gln Val Glu Ile His Arg Asn Gln Thr Leu LeuPhe Ala Val Leu 290 295 300 Ala Glu Gly Thr Pro Ala Glu Ala Lys Lys LeuLeu Ala Pro Ile Leu 305 310 315 320 Ser Val Gly Ser Pro Gln Val Ser ValGln Val Gly Asn Trp Gly Glu 325 330 335 Val Tyr Ser Gly Phe Gln Ile ProThr Ala Asn Glu Pro Ala Asn Trp 340 345 350 Lys Phe Tyr Ser Gln Phe ThrArg Lys Pro Phe Pro Gly Lys Ala Ile 355 360 365 Asp Val Ile Cys Ser PheMet Lys His Ala Pro Thr Asp Asp Ser Asn 370 375 380 Phe Phe Thr Gln AlaPhe Gly Gly Ala Val Arg Arg Ser Pro Arg Gly 385 390 395 400 Gly Thr AlaPhe Pro His Arg Asp Ala Leu Phe Tyr Ser Glu Pro Gly 405 410 415 Ala GlyTrp Gly Thr Arg Ser Asp Gln Pro Gly Ile Cys Asp Pro Leu 420 425 430 ThrPro Gln Ala Gln Ala Trp Ile Ala Glu Phe Ser Gln Ala Leu Arg 435 440 445Pro Tyr Val Asp Gly Ala Tyr Val Asn Val Pro Asn Val Gly Met Gln 450 455460 Asp Trp Glu Thr Ala Tyr Trp Arg Gly Asn Phe Asp Arg Leu Arg Glu 465470 475 480 Ile Lys Ala Lys Tyr Asp Pro His Asn Val Phe Gln Tyr Asp GlnSer 485 490 495 Ile Pro Pro Ala Ser Arg 500 4 1237 PRT Streptomycesatroolivaceus 4 Met Pro Lys Leu Pro Ile Pro Arg Pro Gly Pro Ile Ala LeuAla Val 1 5 10 15 Ala Ala Ala Leu Thr Leu Gly Val Ile Pro Ala Thr AlaAla Val Ser 20 25 30 Ala Tyr Ser Ser Gly Glu Val Ser His Asn Asp Ala GlyHis Arg Thr 35 40 45 Pro Gly Asp Ala Gly Thr His Thr Val Thr Leu Ile ThrGly Asp Lys 50 55 60 Val Thr Ile Gly Thr Ala Ala Asp Gly Thr Val Val ArgSer Phe Arg 65 70 75 80 Ala Ala Ser Gly Ala Thr Thr Gly Phe His Arg AlaVal Val Asp Gly 85 90 95 Ala Thr Tyr Val Tyr Pro Asp Ala Ala Leu Pro TyrVal Ser Ala Gly 100 105 110 Lys Leu Asp Lys Gln Leu Phe Asn Val Thr ArgLeu Ile Thr Glu Gly 115 120 125 Tyr Asp Asp Ala His Ser Ser Arg Leu ProLeu Ile Val Arg Tyr Thr 130 135 140 Gly Ala Ala Ala Lys Ala Arg Thr ArgThr Lys Met Ala Gly Ser Thr 145 150 155 160 Asp Val Arg Arg Leu Asp SerIle Gln Gly Ala Ala Leu Ala Gln Asn 165 170 175 His Lys Gln Ala Pro GluPhe Trp Ser Ser Leu Thr Gly Gly Ser Gly 180 185 190 Ala Ala Ala Arg SerAla Lys Pro Ser Phe Ala Gly Gly Val Ala His 195 200 205 Val Trp Leu AspGly Arg Val Lys Ala Asp Leu Ala Asp Ser Thr Ala 210 215 220 Gln Ile GlyAla Gln Lys Val Trp Ala Glu Gly Asp Thr Gly Gln Asp 225 230 235 240 ValLys Val Ala Met Leu Asp Ser Gly Ala Asp Thr Glu His Pro Asp 245 250 255Leu Val Gly Gln Val Ser Asp Ser Ala Ser Phe Val Pro Gly Glu Asp 260 265270 Asp Ile Ala Asp Tyr Asn Gly His Gly Thr His Val Ala Ser Thr Ile 275280 285 Val Gly Thr Gly Ser Ala Ser Asp Gly Lys Glu Arg Gly Val Ala Ser290 295 300 Gly Ala Arg Leu Ser Val Gly Lys Val Leu Asn Ser Glu Gly SerGly 305 310 315 320 Gln Glu Ser Trp Ile Ile Ala Gly Met Glu Trp Ala AlaArg Asp Gln 325 330 335 Lys Ala Arg Ile Ile Ser Met Ser Leu Gly Gly GlyGly Asp Lys Asn 340 345 350 Asp Pro Met Ser Gln Ala Val Asp Glu Leu SerHis Asp Thr Gly Ala 355 360 365 Leu Phe Val Ile Ala Ala Gly Asn Gly GlyPro His Ser Ile Ser Ser 370 375 380 Pro Gly Ala Ala Asp Ser Ala Leu ThrVal Gly Ala Val Asp Ser Thr 385 390 395 400 Asp Thr Leu Ala Asp Phe SerSer Gln Gly Pro Arg Asp Gly Asp Gly 405 410 415 Gly Leu Lys Pro Glu IleThr Ala Pro Gly Val Asp Ile Val Ala Ala 420 425 430 Arg Ser His Tyr LysArg Gly Ser Gly Tyr Tyr Thr Thr Met Ser Gly 435 440 445 Thr Ser Met AlaThr Pro His Val Ala Gly Val Ala Ala Leu Leu Ala 450 455 460 Ala Glu HisPro Asp Trp Thr Gly Thr Gln Leu Lys Glu Ala Leu Val 465 470 475 480 SerSer Ala Lys Ala Thr Pro Ala Tyr Thr Pro Tyr Gln Ala Gly Ala 485 490 495Gly Arg Leu Asp Ala Pro Ala Ala Val His Thr Thr Val Phe Ala Thr 500 505510 Thr Thr Ala Tyr Ser Gly Phe His Thr Trp Pro Pro Lys Pro Gly Glu 515520 525 Thr Asp Val Arg Thr Val Thr Tyr Thr Asn Val Gly Asp Ala Pro Val530 535 540 Ser Leu Asn Leu Ala Val Asn Gly Thr Val Pro Ala Gly Leu PheSer 545 550 555 560 Leu Ser Glu Asp His Val Thr Val Pro Ala His Gly ThrAla Thr Val 565 570 575 Thr Leu Thr Ala Ala Leu Asp Lys Leu Ala Gly AspGln Ser Val Ser 580 585 590 Ala Val Ile Thr Gly Thr Asp Gly Ser Gly AlaVal Arg Ala Arg Thr 595 600 605 Leu Ile Gly Ala Ala Arg Glu Gly Gln ArgGln Asn Leu Thr Val Val 610 615 620 Ala Lys Asp Arg Ala Gly Lys Pro LeuAla Gly Arg Val Val Leu Thr 625 630 635 640 Ala Glu His Leu Phe Thr ValMet Asp Leu Asp Ala Ser Gly Thr Gly 645 650 655 Thr Thr Arg Leu Pro ValGly Ser Tyr Ser Gly Trp Leu Thr Gly Asp 660 665 670 Val Gln Gly Ala GluGly Pro Asn Ser Leu Gly Met Val Leu Leu Ala 675 680 685 Phe Asn Asp ValLys Leu Asp Gln Asp Arg Thr Val Thr Leu Asp Gly 690 695 700 Arg Lys AlaArg Arg Ile Leu Ala Arg Val Pro Gln Gln Thr Thr Ala 705 710 715 720 ValAla Pro Arg Leu Asp Ala His Arg Ser Phe Thr Asp Ser Leu Val 725 730 735Glu Thr Ser Met Leu Pro Asn Ser Ser Tyr Asp Ser Ile Trp Ala Leu 740 745750 Pro Thr Gly Lys Lys Val Thr Asp Gly Glu Phe Glu Ala Gly Ala Arg 755760 765 Phe Arg Leu Glu Gln Pro Ala Leu Thr Leu Asn Thr Lys Ser Thr Thr770 775 780 Phe Asn Asp Pro Leu Val Lys Arg Ala Ala Thr Pro Leu Pro AlaGly 785 790 795 800 Ser Arg Asn Leu Thr Ala Val Phe Ala Gly Asp Gly ThrAla Glu Glu 805 810 815 Leu Ala Arg Gln Asn Val Arg Gly Lys Ala Val ValVal Arg Gly Ala 820 825 830 Gly Thr Glu Gly Ile Lys Ala Gln Ala Glu AlaAla Ala Ala Ala Gly 835 840 845 Ala Arg Leu Leu Leu Val Val Asn Asp GlyVal Gly Arg Leu Gln Pro 850 855 860 Trp Val Glu Asn Pro Trp Ser Pro GlnSer Pro Ala Pro Val Thr Val 865 870 875 880 Ala Thr Leu Thr Ala Asp GlnGly Ala Glu Leu Ile Gly Ser Leu Arg 885 890 895 His Gly Ala Val Ser LeuLys Val Thr Ser His Pro Thr Thr Asp Tyr 900 905 910 Val Tyr Asp Val ValHis His Trp Thr Gly Ala Val Pro Ala Asp Pro 915 920 925 Thr Trp Arg GluGlu Ser Lys Asp Leu Gly Arg Val Asn Val Ser Phe 930 935 940 Arg Asn TyrArg Gln Gly Lys Ala Met Glu Phe Arg Pro Asp Val Trp 945 950 955 960 ArgGly Trp Ala Val Gly Asn Gln Leu Thr Ala Pro Ala Gln Gly Glu 965 970 975Arg Thr Asp Trp Val Thr Ser Gly Thr Thr Trp Leu Asp Asp Ala Phe 980 985990 Ile Gln Gly Glu Thr Gly Gln His Ser Ile Asp Pro Leu Gln Tyr Ser 9951000 1005 Ala Arg Lys Thr Gly Lys Val Asn Trp Phe Gly Pro Ile Gln Arg1010 1015 1020 Pro Arg Met Gly Pro Val Asn Tyr Gln Pro Val Arg Tyr LeuAsp 1025 1030 1035 Thr Met Tyr Ile Thr Ala Pro Gly Trp Gly Asp Ser GlySer Gly 1040 1045 1050 His Val Gly Glu Ala Gly Ala Asn Phe Asp Val LysAsp Trp Met 1055 1060 1065 Ser Leu Tyr Gln Gly Asp Gln Gln Leu Asp TrpGly Asn Ala Glu 1070 1075 1080 Tyr Leu Gln Val Pro Gly Leu Gly Ala GluArg Leu Pro Tyr Arg 1085 1090 1095 Leu Val Leu Asp Asn Asn Arg Gly AlaTrp Ala Asn Pro Tyr Ser 1100 1105 1110 Thr His Thr Leu Thr Glu Trp AsnPhe Thr Ser Ala Val Thr Ser 1115 1120 1125 Ser Gln Ser Ala Val Ser LeuPro Leu Ile Gln Leu Asp Tyr Gly 1130 1135 1140 Val Asp Thr Asp Lys SerGly Arg Ala Asp Arg His Ala Gly Leu 1145 1150 1155 Thr Val Thr Ala SerHis Leu Pro Gly Thr Thr Ala Ala Ile Ala 1160 1165 1170 Lys Pro Ser ValGlu Val Ser Tyr Asp Asp Gly Ala Thr Trp Gln 1175 1180 1185 Arg Thr GlyLeu Ser Arg Cys Gly Asp Gly Trp Arg Thr Asn Leu 1190 1195 1200 Arg AlaPro Arg Ser Ala Gly Phe Ala Thr Leu Arg Val Gly Ala 1205 1210 1215 ArgAsp Asn Ala Gly Asn Thr Val Ser Gln Thr Ile Thr Arg Ala 1220 1225 1230Phe Gly Leu Arg 1235 5 262 PRT Streptomyces atroolivaceus 5 Met Ala AspGlu Pro Ala Ser Pro Ala Val Gly Arg Leu Arg His Pro 1 5 10 15 Pro GlyArg Arg Pro Gly Gln Arg Gly Gln His Arg Leu Ala Asp Asp 20 25 30 His ProGly Leu Arg Pro Ala Leu Thr Ser Gln Pro Gly Arg Ser Pro 35 40 45 Gly ThrAla Pro Cys Gly Trp Asp Arg Ser Arg Arg Pro His Pro His 50 55 60 Gly IlePro Ser Arg Gln Arg Ile Tyr Glu Asp Thr Pro Ser Arg Asn 65 70 75 80 GlyLys Arg Val Leu Val Asp Arg Leu Trp Pro Arg Gly Met Ser Lys 85 90 95 AlaArg Ala His Leu Asp Glu Trp Ile Arg Asp Val Ala Pro Pro Ala 100 105 110Ala Gly Arg Pro Ser Ala Ser Leu Leu Pro Glu Thr Trp Arg Pro Thr 115 120125 Arg Ala Arg Arg Val Pro Gly Gln Thr Ile Arg Thr Ala Ala Gly Phe 130135 140 Thr Thr His Ser Ala Cys Thr Arg Ser Gly Ala Ala Gly Glu Gly Val145 150 155 160 Asp Arg Phe Leu Pro Val Val Ser Pro Ala Gly Trp Val GlySer Leu 165 170 175 Gly Ser Ala Glu Arg Glu Arg Arg Arg Ser Ala Leu SerGln Ala Val 180 185 190 Val Ser Asp Ser Met Gly Cys Gly Val Cys Thr LysThr Pro Arg Ser 195 200 205 Gly Ser Arg Pro Thr Leu Trp Pro Tyr Ser ProThr Arg Arg Pro Val 210 215 220 Gly Ser Leu Arg Leu Phe Gly Met Cys ArgAla Ser His Pro Ala Pro 225 230 235 240 Val Thr Ser Lys Pro Pro Ala ProIle Leu Trp Ala Ser Val Ser Glu 245 250 255 Ala Arg Arg Thr Glu Arg 2606 401 PRT Streptomyces atroolivaceus 6 Met Ser Ala Arg Gln Glu Thr SerThr Ala Thr Ala His His Val Val 1 5 10 15 Ile Leu Gly Ala Gly Tyr AlaGly Met Ser Ala Ala Ile Gln Leu Ala 20 25 30 Ala Arg Val Lys Gly Arg GluAsp Val Gln Val Thr Val Val Asn Ala 35 40 45 Gln Glu Arg Phe Thr Glu ArgMet Arg Leu His Leu Thr Ala Thr Gly 50 55 60 Gln Arg Leu Ala Glu Leu SerIle Pro Glu Leu Leu Glu Gly Thr Glu 65 70 75 80 Ala Arg Phe Val Arg GlyTrp Val Thr Ala Val Asp Ala Asp Ala Lys 85 90 95 Thr Val Arg Ile Asp AspAsp Arg Val Leu Pro Tyr Asp Thr Leu Val 100 105 110 Tyr Gly Leu Gly GlyVal Ala Asp Thr Cys Thr Val Pro Gly Val Glu 115 120 125 Asp His Ala TyrThr Leu Ser Ser Ala Gln Asp Ala Glu Leu Leu Ala 130 135 140 Val Arg LeuGly Arg Leu Gly Ser Gly Thr Val Val Val Ala Gly Ser 145 150 155 160 GlyLeu Thr Gly Val Glu Ser Ala Ala Glu Ile Ala Glu Arg His Pro 165 170 175Glu Leu Asp Val Val Leu Leu Gly Arg Ala Glu Pro Gly Ala Thr Met 180 185190 Asn Pro Lys Ala Lys Ala Tyr Val Gln Ala Ala Leu Asp Arg Leu Gly 195200 205 Val Arg Val Arg Ala Gly Ala Glu Val Val Lys Val Leu Ser Asp Thr210 215 220 Val Glu Leu Ala Gly Gly Glu Ser Ile Ala Ala Asp Val Val LeuTrp 225 230 235 240 Thr Ser Gly Thr Arg Ala Ser Pro Leu Ala Ala Ala AlaGly Leu Ala 245 250 255 Val Asp Glu Arg Gly Arg Ile Val Thr Asp Ala AlaLeu Arg Ser Val 260 265 270 Ser His Pro Asp Val Tyr Ala Val Gly Asp AlaAla Ala Ile Arg Gln 275 280 285 Gly Tyr Gly Val Met His Gly Thr Cys GlnGly Gly Met Pro Thr Gly 290 295 300 Val His Ala Ala Leu Ser Ile Glu CysVal Leu Lys Gly Lys Gln Pro 305 310 315 320 Lys Pro Phe Arg Phe Gly TyrTyr His Thr Pro Val Ser Leu Gly Arg 325 330 335 Gly Asp Ala Val Val GlnPhe Thr His Pro Asp Asp Ser Pro Arg Arg 340 345 350 Ile Cys Leu Thr GlyArg Met Ala Val Arg Tyr Lys Glu Met Val Thr 355 360 365 Ala Ser Pro TrpPro Thr Tyr Gly Arg Met Lys Lys Met Pro Ala Ser 370 375 380 Gly Ala PheTrp Pro Arg Gly Gly Arg Ser Ala Arg Val Arg Glu Ala 385 390 395 400 Arg7 306 PRT Streptomyces atroolivaceus 7 Met Thr Asp Pro Thr Ala His ProGly Gln Gln Ala Phe His Glu Tyr 1 5 10 15 Arg Lys Leu Leu Phe Ser ValAla Tyr Arg Leu Leu Gly Thr Ala Ala 20 25 30 Asp Ala Glu Asp Val Val GlnAsp Ala Trp Ile Lys Trp Ser Ala Ala 35 40 45 Asp Arg Ser Gln Val Ala AspPro Arg Ala Tyr Leu Thr Arg Ile Val 50 55 60 Ser Asn Leu Ala Leu Glu ArgLeu Arg Ser Thr Arg His Lys Arg Glu 65 70 75 80 Thr Tyr Met Gly Pro TrpLeu Pro Glu Pro Ile Leu Thr Gly Gly Asp 85 90 95 Ala Ser Glu Ala Val ThrGlu Ala Glu Ser Val Ser Met Ala Met Leu 100 105 110 Val Val Leu Glu ThrLeu Ser Pro Leu Glu Arg Ala Val Phe Val Leu 115 120 125 Lys Glu Val PheAsp Phe Ser His Ala Glu Ile Ala Glu Ala Val Glu 130 135 140 Arg Ser GluAla Ala Val Arg Gln Ala Ala His Arg Ala Arg Glu His 145 150 155 160 ValArg Ala Arg Arg Pro Arg Phe Ala Thr Asp Arg Ser Arg Gln Cys 165 170 175Glu Val Thr Glu Arg Phe Phe Ala Ala Ala Thr Gly Gly Asp Ile Asn 180 185190 Thr Leu Met Glu Leu Leu Ser Pro Asp Val Thr Leu Trp Thr Asp Gly 195200 205 Gly Gly Lys Val Arg Gln Ala Met Arg Pro Val Val Gly Ala Ala Thr210 215 220 Val Ala Ala Trp Phe Ala Ala Ile Gly Thr Val Ala Tyr Gln GlyVal 225 230 235 240 Glu Pro Ala Asp Met Lys Ala Glu Leu Val Glu Ile AsnGly Gly Pro 245 250 255 Gly Met Val Phe Ser Gly Leu Gly Arg Val Ile AlaThr Val Thr Phe 260 265 270 Asp Phe Asp Ala Asn Gly Arg Ile Thr Ala IleHis Asn Val Ala Asn 275 280 285 Pro Asp Lys Leu Gln Ala Ile Ala Asp GlyThr Ala Arg Asp Val Gly 290 295 300 Thr Arg 305 8 327 PRT Streptomycesatroolivaceus 8 Met Val Ala Asn Glu Gly Val Ala Val Val Gly Ala Asp ValAsp Asp 1 5 10 15 Arg Gly Trp Ser Asp Thr Arg Ala Trp Val Glu Lys ArgVal Gly Gly 20 25 30 Gly Cys Arg Ile Asp Ala Ala Glu Arg Leu Arg Gly GlyTrp Thr Ser 35 40 45 Glu Met Arg Arg Leu Arg Thr Gly Gly Thr Gly Glu ProGln Trp Met 50 55 60 Val Leu Arg Ser Phe Val Lys Pro Phe Phe Val Gln HisAla Lys Gly 65 70 75 80 Leu Leu Thr Arg Glu Ala Glu Ile Leu Arg Leu LeuSer Glu Thr Asp 85 90 95 Val Pro Val Ala Glu Phe His Gly Val Asp Ala ThrAla Val Tyr Cys 100 105 110 Asp His Pro Ser Leu Leu Met Ser Leu Leu ProGly Ser Val Arg Leu 115 120 125 Gly Glu Glu Glu Ala Gly Leu Arg Ser GlnLeu Leu Ala Arg Gln Leu 130 135 140 Val Gly Ile His Gln Val Arg Val ProVal Glu Ala Arg Pro Arg Val 145 150 155 160 Tyr Gln Ala Trp Thr Ser ProAsp Arg Val Arg Leu Pro Glu Ser Thr 165 170 175 Glu Arg Leu Asp Leu TrpGln Arg Ala Val Asp Ile Ile Arg Arg Asp 180 185 190 Pro Pro Asp Tyr ArgPro Cys Phe Leu His Arg Asp Phe His Pro Gly 195 200 205 Asn Val Leu PheThr Gly Gln Gly Ala Arg Ser Glu Ile Thr Gly Val 210 215 220 Val Asp TrpVal Glu Ala Ser Trp Gly Pro Ala Asp Leu Asp Val Ala 225 230 235 240 HisCys Ser Thr Ala Leu Ala Leu Leu His Gly Ala Glu Glu Gly Met 245 250 255Arg Leu Ala Asp Leu Tyr Leu Ala Ser Gly Gly Thr Leu Ser Glu Asp 260 265270 Pro Ala Ala His Leu Tyr Trp Arg Val Leu Asp Ala Leu Ala Phe Ala 275280 285 Pro Asp Ala Glu Lys Val Ala Val Pro Trp Arg Glu Leu Gly Arg Lys290 295 300 Asp Leu Thr Ser Asp Leu Leu Thr Ser Arg Leu Glu Glu Tyr LeuGlu 305 310 315 320 Glu Leu Phe Arg Arg Phe Gly 325 9 198 PRTStreptomyces atroolivaceus 9 Met Gly Arg Val Ser Gln Ala Gln Ala Gln GluAsn Arg Gln Arg Val 1 5 10 15 Val Ala Thr Ala Ser Arg Met Phe Arg GluLys Gly Thr Ala Val Ser 20 25 30 Val Ala Asp Leu Met Lys Ala Ala Gly LeuThr His Gly Gly Phe His 35 40 45 Lys Gln Phe Ala Ser Lys Asp Asp Leu ValAsp Glu Ala Ile Ala His 50 55 60 Ala Phe Ala Glu His Ala Ala His Ser AlaAla Thr Phe Glu Glu Arg 65 70 75 80 Ala Gly Glu His Glu Ala Ala Arg ArgThr Leu Ile Glu Gln Tyr Leu 85 90 95 Ser Val Trp His Arg Asp His Ala AlaAsp Gly Cys Pro Val Ser Gly 100 105 110 Phe Ala Ala Asp Leu Gly Arg GluPro Asp Gln Ala Ala Arg Ala Arg 115 120 125 His Val Tyr Ile Asn Gly IleArg Asn Arg Ala Ala Arg Leu Ala Thr 130 135 140 Gly Asp Asp Asp Gly MetAla Gln Leu Cys Thr Met Val Gly Ala Leu 145 150 155 160 Ala Leu Ala ArgAla Thr Arg Gly Asp Pro Ile Ser Glu Glu Leu Leu 165 170 175 Gln Ala AlaArg Thr Ala Leu Ala Glu Ser Gly Thr Lys Gln Ser Glu 180 185 190 Pro GlnGln Arg Met Asp 195 10 538 PRT Streptomyces atroolivaceus 10 Met Ala SerPro Pro Thr Pro Ser Glu Ser Ser Thr Pro Thr Arg Ser 1 5 10 15 Thr ProPro Val Pro Pro Ala Gln Arg Arg Arg Val Ser Glu Ala Val 20 25 30 His HisArg Arg Trp Ala Ile Leu Ala Val Leu Ala Phe Ser Leu Leu 35 40 45 Val ValMet Leu Asp Asn Ser Ile Leu Asn Val Ala Met Lys Thr Ile 50 55 60 Ala GlnPro Ala Pro Thr Gly Leu Gly Ser Thr Gln Ser Gln Leu Glu 65 70 75 80 TrpAla Val Asn Ser Tyr Thr Leu Val Phe Ala Gly Leu Leu Phe Thr 85 90 95 AlaGly Leu Leu Gly Asp Arg Leu Gly Arg Lys Lys Val Leu Leu Phe 100 105 110Gly Met Leu Val Phe Gly Ala Gly Ser Ala Leu Ser Ala Leu Ser Asp 115 120125 Ser Ser Gly Gln Leu Ile Thr Phe Arg Ala Val Met Gly Leu Gly Gly 130135 140 Ala Phe Ile Leu Pro Ala Thr Leu Ala Ile Ile Met Asn Val Phe Glu145 150 155 160 Arg Glu Glu Gln Pro Lys Ala Ile Gly Ile Trp Thr Gly ValVal Gly 165 170 175 Phe Ala Ile Ala Ala Gly Pro Ile Ala Gly Gly Val LeuLeu Glu His 180 185 190 Phe Trp Trp Gly Ser Val Phe Leu Val Asn Val ProIle Val Val Ala 195 200 205 Ala Ile Ala Ala Met Ala Ala Ile Val Pro AspSer Lys Asp Pro Arg 210 215 220 Pro Ser Arg Leu Asp Ala Val Gly Val LeuLeu Ser Ile Val Gly Leu 225 230 235 240 Ala Val Leu Val Tyr Gly Ile IleLys Gly Gly Gln Leu Gly Asp Phe 245 250 255 Thr Arg Pro Glu Ala Trp AlaThr Ile Leu Gly Gly Leu Ala Val Leu 260 265 270 Ala Gly Phe Val Val TyrGlu Ala His Ser Asp His Pro Ala Leu Asp 275 280 285 Val Thr Tyr Phe ArgAsn Arg Arg Phe Ser Ala Ser Val Ala Ala Ile 290 295 300 Gly Leu Val PhePhe Ala Leu Met Gly Val Thr Phe Phe Ser Val Phe 305 310 315 320 Tyr AsnGln Ser Val Arg Gly Tyr Ser Val Leu Gln Ser Gly Leu Leu 325 330 335 ValLeu Pro Leu Ala Leu Ser Gln Met Phe Phe Ala Pro Arg Ala Arg 340 345 350Leu Val Val Asp Arg Tyr Gly Ala Arg Ala Val Cys Ala Gly Gly Leu 355 360365 Ala Leu Thr Ala Val Ala Phe Thr Gly Phe Leu Leu Leu Asp Gln Asp 370375 380 Thr Pro Ile Trp Val Leu Glu Val Leu Phe Phe Leu Met Gly Thr Gly385 390 395 400 Met Ala His Val Met Pro Pro Ala Thr Val Met Ile Met SerSer Leu 405 410 415 Pro Arg Glu Lys Ala Gly Ser Gly Ser Ala Val Asn AsnThr Phe Arg 420 425 430 Gln Val Gly Gly Ala Ile Gly Val Ala Val Leu GlySer Leu Leu Ser 435 440 445 Thr Thr Tyr Arg Asn Gly Ile Asp Gly His LeuGly Thr Leu Pro Ala 450 455 460 Ala Asp Arg His Ala Ala Gly Glu Ser ValGln Ala Thr Leu Ala Val 465 470 475 480 Ala Glu Lys Leu Gly Pro Ala GlySer Ala Leu Val Arg Pro Ala His 485 490 495 Asp Ala Phe Ile His Ala MetHis Ile Thr Ala Ala Gly Ser Ala Ala 500 505 510 Val Ala Leu Val Gly ThrVal Val Val Leu Val Phe Leu Pro Gly Arg 515 520 525 Lys Val Thr Ala AlaThr Gly Gly Asp Lys 530 535 11 300 PRT Streptomyces atroolivaceus 11 MetThr Ala Asp Asn Leu Val Leu Ile Pro Gly Ala Gly Gly Val Gly 1 5 10 15Arg Thr Val Phe Glu His Leu Arg Ala Gln Asp Val Pro Val Arg Phe 20 25 30Met Val Arg Arg Asp Asp Glu Arg Ala Ala Glu Leu Arg Ala Leu Gly 35 40 45Ala Glu Val Val Ile Gly Asp Leu Thr Arg Pro Glu Thr Val Ala Ala 50 55 60Ala Leu Gln Gly Val Ala Arg Met His Phe Gly Ile Ser Val Ser Pro 65 70 7580 Asp Gln Leu Leu Ala Ala Thr Val Val Ala Ser Val Ala Arg Glu Tyr 85 9095 Gly Glu Leu Glu Ala Leu Val Asp Leu Ser Gln Met Thr Val Ser Gln 100105 110 Met Thr Ala Thr Ser Thr Glu Glu Ser His Gln Gln Arg Leu His Trp115 120 125 Leu Thr Glu Gln Met Leu Asp Trp Ser Gly Leu Pro Val Val HisVal 130 135 140 Arg Pro Thr Ala Phe Leu Asp Asn Pro Leu Phe Thr Thr IleAla Ala 145 150 155 160 Arg Ser Ile Gln Ala Asn Gly Thr Ile Ala Leu ProPhe Gly Thr Gly 165 170 175 Arg Thr Ser Pro Val Ala Ala Asp Asp Val AlaArg Thr Ile Ala Thr 180 185 190 Val Leu Arg Asp Pro Ala Pro His Ile GlyGln Val Tyr Glu Leu Thr 195 200 205 Gly Pro Arg Ser Val Asp Met Thr GluLeu Ala Ala Glu Phe Ser Arg 210 215 220 Ala Leu Asn Arg Pro Val Ser TyrVal Asp Leu Pro Leu Asp Arg Trp 225 230 235 240 Glu Ala Gln Leu Pro LysLeu Gly Met Ser Pro His Val Ala Gln His 245 250 255 Val Ala Thr Met AlaArg Leu His Arg Asp Asn Arg Tyr Asp Arg Thr 260 265 270 Ala Asp Gly ValGlu Arg Val Thr Gly Thr Pro Ala Met Ser Ile Glu 275 280 285 Ala Phe ValAla Ala Arg Arg Asp Leu Tyr Leu Gly 290 295 300 12 197 PRT Streptomycesatroolivaceus 12 Met Thr Leu Asp Asp Leu Leu Leu Pro Asp Thr Pro Val CysThr Ala 1 5 10 15 Ala Thr Asp Val Ala Arg Ala Tyr Cys Ser Pro Ala LeuLeu Asn His 20 25 30 Cys Val Arg Ser Tyr Ile Trp Ala Ala Val His Gly AlaGlu Gln Asp 35 40 45 Ile Ala Phe Asp Pro Glu Leu Leu Tyr Val Gly Ala MetPhe His Asp 50 55 60 Ile Ala Leu Val Pro Glu Phe Asp Ser His Thr Val ProPhe Asp Asp 65 70 75 80 Ala Ser Ala His Val Ala Arg Val Phe Ala Ala GlyAla Gly Trp Ser 85 90 95 Glu Ser Arg Arg Glu Arg Leu Ala Glu Val Val IleSer His Met Leu 100 105 110 Asp Glu Val Asp Val Thr Lys Asp Pro Glu GlyHis Leu Leu Glu Arg 115 120 125 Ser Thr Ser Leu Asp Ile Ser Gly Arg HisMet Asp Asp Phe Ser Ala 130 135 140 Ala Phe Lys Ala Glu Val Leu Ala ArgHis Pro Arg Ala Gly Ile Ala 145 150 155 160 Glu Glu Phe Leu Ala Cys PheArg Asp Gln Ala Glu Arg Lys Pro Gly 165 170 175 Ser Ser Pro Ala Thr AlaLeu Arg Asp Gly Ile Ala Glu Arg Ile Leu 180 185 190 Thr Asn Ser Leu Asp195 13 353 PRT Streptomyces atroolivaceus 13 Met Thr Thr Leu Thr Phe AlaGlu Ala Ser Ala His Ser Gly Gly Asp 1 5 10 15 Leu Arg Tyr Leu Pro AlaGly His Gly Glu Val Leu Asp Leu Ser Thr 20 25 30 Cys Val Thr Arg Ser GlyPro Ser Pro Arg Ala Leu Ala Ala Ala Ala 35 40 45 Ala Phe Asp Pro Ser SerLeu Thr Val His Pro Tyr Gly Ala Asp Asp 50 55 60 Arg Phe Arg Thr Thr TyrAla Ala Tyr Leu Gly Val Asp Ala Arg Gln 65 70 75 80 Leu Ala Val Thr ArgGly Ile Ser Glu Pro Leu Ser Val Leu Ala Ser 85 90 95 Leu Leu Pro Ala GluArg Ser Ala Ala Leu Thr Pro Asp Tyr Thr Gly 100 105 110 Thr Ile Arg ArgTrp Ala Arg His Leu Pro Pro Ala Pro Asp Glu Arg 115 120 125 Asp Cys ValAsp Ser Arg Val Arg Arg Leu Asp Ala Ala Met Arg His 130 135 140 Tyr AlaTyr Val Val Phe Ser Asn Pro Asn Asn Pro Leu Gly Leu Thr 145 150 155 160Val Pro Arg Asp Val Leu Ala Glu Leu Leu Ser Arg His Pro Arg Cys 165 170175 Thr Leu Val Val Asp Glu Ala Tyr Val Asp Tyr Met Gly Asp Arg Arg 180185 190 Gln Ser Ala Met Thr Leu Pro His Arg Asn Leu Ala Val Leu Ala Ser195 200 205 Pro Gly Lys Pro Leu Gly Ile Ala Gly Thr Arg Thr Gly Val LeuTrp 210 215 220 Thr Pro Asp Asp Val Leu Arg Glu Arg Val Ala Arg Arg LeuPro Glu 225 230 235 240 Trp Pro Leu Ser Ala Tyr Asp Ala His Val Ala CysAla Ala Leu Gln 245 250 255 Asp Thr Ala Trp Ile Ala Ser Ala Leu Glu GlyAla Arg Glu Asp Ala 260 265 270 Arg Arg Leu Glu Ala Val Leu Val Arg CysPhe Gly Asp Ala Val Val 275 280 285 Thr Gly Val Pro Val His Tyr Arg PheVal Tyr Asp Glu Arg Pro Glu 290 295 300 Arg Leu His Ala His Leu Leu AlaAsp Gly Ile Ala Thr Arg Leu Leu 305 310 315 320 Val Asp Asp Glu Pro GlyArg Val Arg Gly Leu Arg Val Leu Ala Pro 325 330 335 Gly Ser Gly Glu LeuAla Arg Leu Glu Ala Ser Leu Asn Ala Phe Val 340 345 350 Arg 14 774 PRTStreptomyces atroolivaceus 14 Met Glu Phe Ser Ser Arg Ser Ser Thr SerSer Thr Thr Gly Ala Pro 1 5 10 15 Leu Leu Ser Val Arg Ser Ala Gly GluIle Val Pro Val Leu Cys Pro 20 25 30 Asp Gly Phe Glu Val Gly Gly Gly ProArg Val Val Glu Val Val Ala 35 40 45 Asp Arg Trp Ala Gly Lys Ser Thr ValLeu Asp Gly Leu Ser Ala Ala 50 55 60 Leu Ala Gly Arg Gly Trp Thr Val AlaAla Gly Ala Ala Ser Pro Ala 65 70 75 80 Arg Ser Arg Ala Pro Phe Gly ValPhe Val Glu Ala Leu Glu Glu Leu 85 90 95 Val Glu His Leu Asp Pro Asp ArgLeu Gly Gly Arg Gly Arg Asp His 100 105 110 Ala Ala Gln Leu Ala Ala LeuPhe Pro Ala Leu Ser Pro Asp Ala Pro 115 120 125 Arg Asp Thr Ser Val AspThr Tyr Arg Ile Cys Arg Ala Met Arg Ala 130 135 140 Leu Leu Glu Met LeuAla Ala Asp Gly Pro Leu Leu Leu Ala Leu Asp 145 150 155 160 Asp Val HisTrp Met Asp Ala Asp Ser Leu Glu Leu Leu Gly Gln Leu 165 170 175 Leu GluGln Pro Val Ala Gly Pro Val Val Ile Ala Val Ala His Arg 180 185 190 GlyArg Pro Gly Asp Arg Val Leu Asn Gly Leu Leu Ala Arg Cys Ala 195 200 205Ser Arg Gly Gly Cys Leu Arg Val Arg Pro Ala Pro Leu Thr Glu Gln 210 215220 Glu Ala Ile Ala Leu Leu Pro Asp Asp Leu Ser Arg Val His Cys Glu 225230 235 240 Thr Leu Leu Val Glu Ser Gly Gly Ser Pro Gly Leu Leu Arg AlaPhe 245 250 255 Ala Thr Leu Arg Gln Val Pro Gly Leu Thr Gly Ala Trp ThrPro Pro 260 265 270 Leu Pro Ala Asp Leu Leu Ala Asp Val Leu Arg Asp PheArg Ala Leu 275 280 285 Ser Ala Glu Gly Trp Gln Val Val Arg Ala Ala AlaVal Leu His Glu 290 295 300 Pro Tyr Ala Ala Asp Asp Leu Lys Glu Ile AlaHis Val Asp Asp Ala 305 310 315 320 Ala Leu Trp Arg Ala Leu Asp Glu LeuIle His Glu Glu Leu Leu Arg 325 330 335 Pro Asp Glu Leu Pro Trp Gln LeuArg Phe Ala Asn Pro Leu Leu Arg 340 345 350 Val Ala Ala Tyr Gln Ser AlaGly Arg Gly Trp Leu Leu Gly Ala His 355 360 365 Ala Gln Ala Ala Arg IleLeu Ala Arg Pro Gly Tyr Pro Ser Ala His 370 375 380 Met Ala Arg His LeuGlu Arg Gly Ala Ala Leu Arg Asp Glu Ser Gly 385 390 395 400 Ala Arg ValLeu Trp Asp Ala Ala Ala Arg Leu Leu Trp Gln Asp Pro 405 410 415 Ser GlnSer Val Ser Trp Ile Arg Ala Ala Leu Ala Ser Arg Ala Gly 420 425 430 GlyAsp Leu Glu Pro Arg Asp Arg Leu Leu Phe Gly Lys Ala Leu Ala 435 440 445Leu Ala Gly Arg Leu Thr Glu Ser Leu Ala Thr Leu Gly Pro Phe Gly 450 455460 Gln Gly Gly Val Glu Ala Gly Leu Arg Val Glu Arg Val Arg Trp Arg 465470 475 480 Ala His Ala His Arg Leu Leu Gly His Thr Asp Arg Ala Gln AspGlu 485 490 495 Leu Glu Ser Leu Leu Ala Glu Leu Glu Pro Gly Glu Glu GlyLeu Arg 500 505 510 Asp Val Val Arg His Ala Leu Ile Glu Leu Ala Leu HisGly Arg Arg 515 520 525 Pro Val Pro Pro Pro Pro Gly Gly Ser Glu Pro GlyMet Pro Gln Gly 530 535 540 Ala Thr Gly Ala Leu Arg Gly Arg Leu Leu AlaGln Leu Ala Val Ala 545 550 555 560 Ala Cys Arg Gly Gly Glu Gln Glu ArgAla Glu Gln Arg Ala Leu Thr 565 570 575 Ala Arg Gln Leu Leu Asp Ala AlaPro Asp Gly Glu Val Ala Glu Gln 580 585 590 Leu Glu Gly Leu Tyr Trp LeuAla Thr Ala Glu Ala Ala Leu Gly Arg 595 600 605 Pro Ser Ala Ala Cys GlyHis Tyr Glu Arg Gly Leu Arg Ile Ala Glu 610 615 620 Thr Arg Arg Leu IleAsp His Val Pro Leu Phe Ala Met Ala Leu Ser 625 630 635 640 Ala Leu GlnLeu His Thr Gly Asp Gly Ala Gly Ala Ala Arg His Ala 645 650 655 Ala CysAla Glu Thr Val Ala Ala Ala Thr Glu Ser Glu His Leu Arg 660 665 670 GluArg Ala Leu Arg Leu Arg Arg Glu Thr Ala Ser Gly Ala Gly Val 675 680 685Ala Val Leu Pro Gly Gln Arg Gln Pro Ala Asp Ala Ala Ala Val Ala 690 695700 Leu Glu Gly Leu Ser Arg Arg Glu Ser Glu Val Ala Leu Leu Val Ser 705710 715 720 Gly Gly Arg Thr Asn Gln Gln Ile Ala Arg Val Leu Glu Leu SerHis 725 730 735 Lys Thr Val Glu Thr Tyr Leu Gly Arg Ile Phe Gln Lys LeuGln Val 740 745 750 Gln Ser Arg Ala Glu Val Ala Ala Met Val Gly Arg SerAsp Arg Leu 755 760 765 Gly Val Ala Ala Gly Ser 770 15 1105 PRTStreptomyces atroolivaceus 15 Met Gln Thr Gln Leu Ser Gln Gly Ala GlnLeu Ser Ala Glu Gln Arg 1 5 10 15 Lys Leu Leu Asp Gln Leu Leu Asp AlaAsp Gly Ile Asp Arg Val Ala 20 25 30 Gly Lys Val Thr Arg Arg Thr Gly AspLeu Ser Arg Ala Pro Leu Ser 35 40 45 Phe Gly Gln Glu Arg Leu Tyr Phe ValAsp Lys Leu Arg Pro Gly Ser 50 55 60 Pro Met Tyr Val Gly Ser Gly Ala LeuArg Leu Arg Gly Ala Leu Asp 65 70 75 80 Val Glu Val Leu Arg Arg Ser IleAla Leu Leu Val Asp Arg His Glu 85 90 95 Val Leu Arg Thr Gly Val Val GluAsp Gly Asp Gly Val Glu Gln Arg 100 105 110 Ile Arg Pro Ala Gly Thr ValGln Val Glu Leu Pro Val Arg Gly Ala 115 120 125 Ala Ser Gly Glu Val AlaAsp Arg Val Arg Glu Ala Ala Ala Glu Gly 130 135 140 Phe Asp Leu Ser AlaPro Pro Leu Leu Arg Ala Val Leu Leu Glu Ile 145 150 155 160 Thr Asp AlaAla Glu Pro Glu Trp Val Leu Val Leu Ser Val His His 165 170 175 Ile AlaVal Asp Gly Trp Ser Leu Gly Leu Leu Met Gly Glu Met Ser 180 185 190 GluLeu Tyr Ser Ala Leu Val Gln Gly Ala Glu Pro Ser Leu Ala Glu 195 200 205Leu Pro Leu Gln Tyr Ala Asp Phe Ala Ser Trp Gln Arg Gly Trp Leu 210 215220 Glu Gln Gly Ile Ile Ala Asp Gln Leu Ala Tyr Trp Ser Asp Gln Leu 225230 235 240 Ser Gly Val Glu Ala Thr Glu Val Pro Ser Asp Arg Pro Arg ProAla 245 250 255 Glu Arg Ser Tyr Leu Gly Asp Thr Val Pro Leu Ala Leu AspArg Pro 260 265 270 Leu Thr Gln Ser Leu Arg Ala Leu Thr Asp Ser Gly GlnSer Thr Leu 275 280 285 Phe Met Ala Leu Thr Ala Val Trp Ala Ser Val LeuGly Arg Phe Ala 290 295 300 Asp Arg Arg Asp Val Val Val Gly Thr Pro ValAla Gly Arg Thr Arg 305 310 315 320 Ala Glu Leu Glu Ser Val Val Gly PhePhe Val Asn Thr Leu Pro Leu 325 330 335 Arg Val Arg Ile Glu Asp Gly AspThr Phe Arg Thr Leu Leu Leu Arg 340 345 350 Thr Arg Glu Val Cys Thr GlnAla Tyr Ala His Gln Asp Val Pro Phe 355 360 365 Glu Arg Ile Val Ala GluLeu Gln Ser Gly Arg Asp Ala Ser Gly Gln 370 375 380 Thr Ala Ile Ala ArgHis Trp Leu Ala Leu His Asn Thr Pro Ala Pro 385 390 395 400 Asp Leu ThrMet Pro Gly Leu Met Ser Glu Ala Leu Pro Ala Leu Ile 405 410 415 Gly ThrVal Arg Cys Asp Leu Ser Val Gln Leu Ala Pro Asp Gly Gln 420 425 430 GlyGly Leu Ser Gly Arg Leu Glu Tyr Ser Thr Glu Leu Phe Asp Arg 435 440 445Ala Thr Ala Lys Thr Leu Ala Ser Ala Phe Glu Thr Leu Ile Ala Ser 450 455460 Val Ala Ala Ala Pro Asp Ser Pro Leu Asp Glu Leu Asp Met Leu Ala 465470 475 480 Pro Gln Ala Arg Glu His Ile Val Arg Asp Leu Ser Gly Ala AlaArg 485 490 495 Gly Ala Leu Thr Gly Pro Trp Val Pro Glu Arg Phe Ala AlaGln Val 500 505 510 Asp Ala His Pro Asp Thr Val Ala Val Val Glu Ala AspGly Gly Gln 515 520 525 Leu Thr Tyr Gly Gln Leu Asp Val Leu Ala Asn ArgVal Ala His Leu 530 535 540 Leu Thr Ala Arg Gly Val Gly Ala Glu Asp ArgVal Gly Val Leu Leu 545 550 555 560 Pro Gly Gly Gly Ala Leu Ala Ala AlaVal Leu Gly Val Leu Lys Ser 565 570 575 Ala Ala Val Val Leu Pro Leu AspPro Gly Tyr Pro Pro Ala Arg Leu 580 585 590 Ala Gln Leu Val Glu Asp GlyAla Pro Arg Leu Val Leu Ser Gly Ala 595 600 605 Gly Ser Thr Asp Leu LeuPro Asp Leu Asp Val Val Asp Val Thr Gly 610 615 620 Glu Glu Val Ala GlyHis Pro Gly His Arg Pro Pro Ala Val Ala Thr 625 630 635 640 Ala Pro AspAla Gly Ala His Leu Leu Phe Thr Ser Gly Thr Ser Gly 645 650 655 Arg ProLys Gly Val Leu Thr Thr Arg Ala Gly Leu Val Asn Arg Leu 660 665 670 MetGly Met Glu Arg Gln Phe Ser Val Gly Ala Gly Asp Arg Val Leu 675 680 685Cys Lys Ala Pro Leu Gly Phe Asp Val Ala Val Trp Glu Leu Leu Leu 690 695700 Pro Leu Val Thr Gly Ala Thr Val Val Ser Cys Arg Val Gly Gly His 705710 715 720 Arg Asp Leu Glu Tyr Leu Arg Glu Leu Ile Asp Arg His Gly ValSer 725 730 735 Val Cys His Phe Val Pro Ser Leu Leu Glu Glu Phe Ala AsnAla Pro 740 745 750 Ala Gly Ala His Pro Ser Leu Arg Leu Leu Leu Ser GlyGly Glu Glu 755 760 765 Leu Pro Ala Pro Leu Ala Glu Arg Ile Leu Asp ArgPhe Gly His Ala 770 775 780 Glu Phe Val Asn Gln Tyr Gly Pro Thr Glu ThrVal Ile Asp Val Thr 785 790 795 800 Ser Gly Arg Val Ser Gly Pro Val ProHis Arg Val Pro Ile Gly Arg 805 810 815 Pro Val Pro Gly Ala Glu Val TyrVal Leu Asp Ala Ser Met Arg Pro 820 825 830 Gln Pro Val Gly Val Val GlyGlu Leu Tyr Val Gly Gly Val Gln Val 835 840 845 Ala Arg Gly Tyr Val GlyArg Ala Ala Leu Thr Ala Glu Arg Phe Val 850 855 860 Pro His Pro Phe AlaHis Gly Gln Arg Leu Tyr Ala Thr Gly Asp Arg 865 870 875 880 Ala Arg LeuLeu Ala Gly Gly Ser Val Glu Phe Leu Gly Arg Thr Asp 885 890 895 Asp GlnVal Lys Ile Arg Gly Asn Arg Val Glu Pro Ala Glu Val Ala 900 905 910 AlaVal Leu Arg Ala His Pro Gln Val Ser Gly Ser His Val Thr Val 915 920 925Gln Arg Thr Gly Ala Ala Pro Arg Leu Thr Gly Tyr Val Thr Ser Asp 930 935940 Ala Ala Asn Ala Asp Glu Leu Thr Glu Glu Leu Arg Glu Phe Leu Arg 945950 955 960 Glu Arg Val Pro Glu Ala Met Val Pro Ala Gln Leu Phe Val LeuArg 965 970 975 Gln Trp Pro Thr Gly Ala His Gly Lys Ile Asp Val Thr AlaLeu Pro 980 985 990 Gln Pro Ser Gly Glu Arg Pro Gly Ala Gln Arg Glu PheAla Ala Pro 995 1000 1005 Val Thr Asp Val Gln Val Arg Leu Ala Ala LeuCys Thr Glu Leu 1010 1015 1020 Leu Glu Val Glu Arg Ile Gly Leu Thr AspSer Phe Phe Asp Leu 1025 1030 1035 Gly Gly His Ser Leu Leu Ala Ile ArgAla Ile Ser Arg Ile Arg 1040 1045 1050 Lys Glu Phe Gly Val Gly Leu GlnIle Gly Gln Phe Phe Lys Ala 1055 1060 1065 Pro Thr Leu Ala Gly Leu AlaGlu Ala Ile Glu Ala Lys Leu Ala 1070 1075 1080 Ala Ala Asp Gly Lys ProAla Gly Pro Ala Pro Ile Pro Arg Ile 1085 1090 1095 Asp Arg Gly Ala GlnArg Arg 1100 1105 16 1105 PRT Streptomyces atroolivaceus 16 Met Gln ThrGln Leu Ser Gln Gly Ala Gln Leu Ser Ala Glu Gln Arg 1 5 10 15 Lys LeuLeu Asp Gln Leu Leu Asp Ala Asp Gly Ile Asp Arg Val Ala 20 25 30 Gly LysVal Thr Arg Arg Thr Gly Asp Leu Ser Arg Ala Pro Leu Ser 35 40 45 Phe GlyGln Glu Arg Leu Tyr Phe Val Asp Lys Leu Arg Pro Gly Ser 50 55 60 Pro MetTyr Val Gly Ser Gly Ala Leu Arg Leu Arg Gly Ala Leu Asp 65 70 75 80 ValGlu Val Leu Arg Arg Ser Ile Ala Leu Leu Val Asp Arg His Glu 85 90 95 ValLeu Arg Thr Gly Val Val Glu Asp Gly Asp Gly Val Glu Gln Arg 100 105 110Ile Arg Pro Ala Gly Thr Val Gln Val Glu Leu Pro Val Arg Gly Ala 115 120125 Ala Ser Gly Glu Val Ala Asp Arg Val Arg Glu Ala Ala Ala Glu Gly 130135 140 Phe Asp Leu Ser Ala Pro Pro Leu Leu Arg Ala Val Leu Leu Glu Ile145 150 155 160 Thr Asp Ala Ala Glu Pro Glu Trp Val Leu Val Leu Ser ValHis His 165 170 175 Ile Ala Val Asp Gly Trp Ser Leu Gly Leu Leu Met GlyGlu Met Ser 180 185 190 Glu Leu Tyr Ser Ala Leu Val Gln Gly Ala Glu ProSer Leu Ala Glu 195 200 205 Leu Pro Leu Gln Tyr Ala Asp Phe Ala Ser TrpGln Arg Gly Trp Leu 210 215 220 Glu Gln Gly Ile Ile Ala Asp Gln Leu AlaTyr Trp Ser Asp Gln Leu 225 230 235 240 Ser Gly Val Glu Ala Thr Glu ValPro Ser Asp Arg Pro Arg Pro Ala 245 250 255 Glu Arg Ser Tyr Leu Gly AspThr Val Pro Leu Ala Leu Asp Arg Pro 260 265 270 Leu Thr Gln Ser Leu ArgAla Leu Thr Asp Ser Gly Gln Ser Thr Leu 275 280 285 Phe Met Ala Leu ThrAla Val Trp Ala Ser Val Leu Gly Arg Phe Ala 290 295 300 Asp Arg Arg AspVal Val Val Gly Thr Pro Val Ala Gly Arg Thr Arg 305 310 315 320 Ala GluLeu Glu Ser Val Val Gly Phe Phe Val Asn Thr Leu Pro Leu 325 330 335 ArgVal Arg Ile Glu Asp Gly Asp Thr Phe Arg Thr Leu Leu Leu Arg 340 345 350Thr Arg Glu Val Cys Thr Gln Ala Tyr Ala His Gln Asp Val Pro Phe 355 360365 Glu Arg Ile Val Ala Glu Leu Gln Ser Gly Arg Asp Ala Ser Gly Gln 370375 380 Thr Ala Ile Ala Arg His Trp Leu Ala Leu His Asn Thr Pro Ala Pro385 390 395 400 Asp Leu Thr Met Pro Gly Leu Met Ser Glu Ala Leu Pro AlaLeu Ile 405 410 415 Gly Thr Val Arg Cys Asp Leu Ser Val Gln Leu Ala ProAsp Gly Gln 420 425 430 Gly Gly Leu Ser Gly Arg Leu Glu Tyr Ser Thr GluLeu Phe Asp Arg 435 440 445 Ala Thr Ala Lys Thr Leu Ala Ser Ala Phe GluThr Leu Ile Ala Ser 450 455 460 Val Ala Ala Ala Pro Asp Ser Pro Leu AspGlu Leu Asp Met Leu Ala 465 470 475 480 Pro Gln Ala Arg Glu His Ile ValArg Asp Leu Ser Gly Ala Ala Arg 485 490 495 Gly Ala Leu Thr Gly Pro TrpVal Pro Glu Arg Phe Ala Ala Gln Val 500 505 510 Asp Ala His Pro Asp ThrVal Ala Val Val Glu Ala Asp Gly Gly Gln 515 520 525 Leu Thr Tyr Gly GlnLeu Asp Val Leu Ala Asn Arg Val Ala His Leu 530 535 540 Leu Thr Ala ArgGly Val Gly Ala Glu Asp Arg Val Gly Val Leu Leu 545 550 555 560 Pro GlyGly Gly Ala Leu Ala Ala Ala Val Leu Gly Val Leu Lys Ser 565 570 575 AlaAla Val Val Leu Pro Leu Asp Pro Gly Tyr Pro Pro Ala Arg Leu 580 585 590Ala Gln Leu Val Glu Asp Gly Ala Pro Arg Leu Val Leu Ser Gly Ala 595 600605 Gly Ser Thr Asp Leu Leu Pro Asp Leu Asp Val Val Asp Val Thr Gly 610615 620 Glu Glu Val Ala Gly His Pro Gly His Arg Pro Pro Ala Val Ala Thr625 630 635 640 Ala Pro Asp Ala Gly Ala His Leu Leu Phe Thr Ser Gly ThrSer Gly 645 650 655 Arg Pro Lys Gly Val Leu Thr Thr Arg Ala Gly Leu ValAsn Arg Leu 660 665 670 Met Gly Met Glu Arg Gln Phe Ser Val Gly Ala GlyAsp Arg Val Leu 675 680 685 Cys Lys Ala Pro Leu Gly Phe Asp Val Ala ValTrp Glu Leu Leu Leu 690 695 700 Pro Leu Val Thr Gly Ala Thr Val Val SerCys Arg Val Gly Gly His 705 710 715 720 Arg Asp Leu Glu Tyr Leu Arg GluLeu Ile Asp Arg His Gly Val Ser 725 730 735 Val Cys His Phe Val Pro SerLeu Leu Glu Glu Phe Ala Asn Ala Pro 740 745 750 Ala Gly Ala His Pro SerLeu Arg Leu Leu Leu Ser Gly Gly Glu Glu 755 760 765 Leu Pro Ala Pro LeuAla Glu Arg Ile Leu Asp Arg Phe Gly His Ala 770 775 780 Glu Phe Val AsnGln Tyr Gly Pro Thr Glu Thr Val Ile Asp Val Thr 785 790 795 800 Ser GlyArg Val Ser Gly Pro Val Pro His Arg Val Pro Ile Gly Arg 805 810 815 ProVal Pro Gly Ala Glu Val Tyr Val Leu Asp Ala Ser Met Arg Pro 820 825 830Gln Pro Val Gly Val Val Gly Glu Leu Tyr Val Gly Gly Val Gln Val 835 840845 Ala Arg Gly Tyr Val Gly Arg Ala Ala Leu Thr Ala Glu Arg Phe Val 850855 860 Pro His Pro Phe Ala His Gly Gln Arg Leu Tyr Ala Thr Gly Asp Arg865 870 875 880 Ala Arg Leu Leu Ala Gly Gly Ser Val Glu Phe Leu Gly ArgThr Asp 885 890 895 Asp Gln Val Lys Ile Arg Gly Asn Arg Val Glu Pro AlaGlu Val Ala 900 905 910 Ala Val Leu Arg Ala His Pro Gln Val Ser Gly SerHis Val Thr Val 915 920 925 Gln Arg Thr Gly Ala Ala Pro Arg Leu Thr GlyTyr Val Thr Ser Asp 930 935 940 Ala Ala Asn Ala Asp Glu Leu Thr Glu GluLeu Arg Glu Phe Leu Arg 945 950 955 960 Glu Arg Val Pro Glu Ala Met ValPro Ala Gln Leu Phe Val Leu Arg 965 970 975 Gln Trp Pro Thr Gly Ala HisGly Lys Ile Asp Val Thr Ala Leu Pro 980 985 990 Gln Pro Ser Gly Glu ArgPro Gly Ala Gln Arg Glu Phe Ala Ala Pro 995 1000 1005 Val Thr Asp ValGln Val Arg Leu Ala Ala Leu Cys Thr Glu Leu 1010 1015 1020 Leu Glu ValGlu Arg Ile Gly Leu Thr Asp Ser Phe Phe Asp Leu 1025 1030 1035 Gly GlyHis Ser Leu Leu Ala Ile Arg Ala Ile Ser Arg Ile Arg 1040 1045 1050 LysGlu Phe Gly Val Gly Leu Gln Ile Gly Gln Phe Phe Lys Ala 1055 1060 1065Pro Thr Leu Ala Gly Leu Ala Glu Ala Ile Glu Ala Lys Leu Ala 1070 10751080 Ala Ala Asp Gly Lys Pro Ala Gly Pro Ala Pro Ile Pro Arg Ile 10851090 1095 Asp Arg Gly Ala Gln Arg Arg 1100 1105 17 330 PRT Streptomycesatroolivaceus 17 Met Thr Ile Glu Val His Asp Ala Val Ala Val Ser Pro ValTrp Glu 1 5 10 15 Gly Arg Ser Leu Pro Ala Leu Val Arg Cys Glu Ser ProGly Gly Ala 20 25 30 Leu Pro Ala Trp Val Ser Ala His Arg Asp Glu Val AspAla Leu Ala 35 40 45 Arg Lys Ala Gly Ala Val Leu Leu Arg Gly Phe Asp ValArg Gly Ala 50 55 60 Gln Asp Phe Arg Ala Val Met Asp Ala Leu Ser Pro GlnVal Leu Ala 65 70 75 80 Tyr Gly Glu Arg Ser Ser Pro Arg Ser Gln Val SerAsp Gly Val Tyr 85 90 95 Thr Ser Thr Glu Tyr Pro Ala Asp Gln Pro Ile LeuLeu His Asn Glu 100 105 110 Gln Ser Tyr Thr Ala Lys Trp Pro Thr Arg IleVal Phe Phe Cys Glu 115 120 125 Arg Ala Ala Thr Glu Gly Gly Arg Thr ProLeu Ala Asp Ser Arg Arg 130 135 140 Ile Leu Ala Arg Leu Arg Pro Glu ThrVal Asp Arg Phe Glu Arg Leu 145 150 155 160 Gly Val Arg Tyr Val Arg AsnTyr Leu Pro Gly Ile Ser Leu Arg Trp 165 170 175 Gln Glu Ala Phe Gln ThrAsp Arg Val Glu Asp Val Ala Ala Tyr Cys 180 185 190 Ala Arg Ala Asp IleThr Pro Glu Trp Val Asp Glu Asp His Leu Arg 195 200 205 Thr Val Gln ValArg Pro Ala Val Arg Arg His Pro Val Thr Gly Glu 210 215 220 Arg Ser TrpPhe Asn His Ala Leu Phe Phe His Val Thr Ser Leu Asp 225 230 235 240 ProGlu Val Ser Ala Gly Leu Leu Glu Ala Leu Asp Glu Glu Asp Leu 245 250 255Pro Tyr Asn Thr Tyr Tyr Gly Asp Gly Ser Pro Ile Glu Ser Glu Thr 260 265270 Leu Ala Glu Leu Arg Ala Ala Tyr Ala Ala Glu Thr Thr Gly Phe Asp 275280 285 Trp Gln Pro Gly Asp Val Leu Val Val Glu Asn Met Leu Val Ala His290 295 300 Ala Arg Glu Pro Phe Val Gly Pro Arg Arg Ile Leu Thr Ala MetAla 305 310 315 320 Asp Ala Ile Asp Ala Gly Glu Val Pro Ala 325 330 18335 PRT Streptomyces atroolivaceus 18 Met Thr Leu Thr Asp Leu Thr ProAla Leu Glu Asp Gly Thr Val Thr 1 5 10 15 Val Arg Pro Leu His Glu GluPro Phe Leu Leu Glu Ile Glu Ala Arg 20 25 30 Glu Pro Gly Leu Asp Leu ProArg Trp Val Ala Gly Arg Lys Glu Glu 35 40 45 Leu Arg Ala Gly Leu Asp ArgCys Gly Ala Leu Leu Leu Arg Gly Phe 50 55 60 Asp Val Pro Asp Pro Gln GluPhe Gly Arg Ala Ala Arg Ala Phe Ser 65 70 75 80 Pro Asp Leu Leu Gly TyrLeu Glu Arg Ala Ala Thr Arg Thr Glu Val 85 90 95 Ala Asp Lys Val Phe ThrSer Thr Glu Leu Gly Glu Asp Gln Trp Ile 100 105 110 Pro Phe His His GluMet Ser Tyr Ser His Asn Trp Pro Gly Leu Leu 115 120 125 Tyr Phe Phe GlyAsp Leu Pro Ser Pro Glu Gly Gly Ala Thr Pro Val 130 135 140 Ala Ser GluArg Arg Val Phe Pro Leu Ile Pro Ala Glu Val Arg Glu 145 150 155 160 ArgPhe Thr Arg His Gly Val Arg Tyr Val Arg Asn Tyr Gly Pro Asp 165 170 175Leu Asn Glu Ser Trp Gln Val Ala Phe Gln Thr Thr Asp Arg Ala Glu 180 185190 Val Glu Ala Tyr Cys Arg Gln Ser Asp Thr Ala Phe Glu Trp Thr Gly 195200 205 Asp Asp Glu Leu Arg Thr Thr Ala Val Arg Gln Ala Val Ala Arg His210 215 220 Pro Arg Thr Gly Glu Thr Val Trp Phe Asn His Ala His Leu PheHis 225 230 235 240 Leu Ser Asn Val Glu Pro Glu Val Ala Glu Ile Leu ValGlu Glu Tyr 245 250 255 Gly Glu Glu Gly Leu Pro Arg Asn Ala Tyr Tyr GlyAsp Gly Ala Pro 260 265 270 Ile Glu Asn Glu Val Ala Glu Leu Ile Arg SerLeu Tyr Arg Glu Ala 275 280 285 Ala Val Ser Phe Thr Trp Arg Arg Gly AspVal Leu Val Val Asp Asn 290 295 300 His Leu Ala Thr His Ala Arg Glu ProPhe Arg Gly Asp Arg Arg Ile 305 310 315 320 Leu Val Ala Met Ser Asp LeuHis Thr Glu Leu Pro Ala Gly Arg 325 330 335 19 313 PRT Streptomycesatroolivaceus 19 Met Leu Leu Arg Pro Thr Thr Asp Ala Asp Leu Ala Arg ValThr Asp 1 5 10 15 Phe Ile Val Glu Glu Pro Ile Ser Trp Ile Pro Ala AspArg Tyr Arg 20 25 30 Ala Glu Leu Ala Glu Gly Met Tyr Arg Pro Gln Trp SerTrp Ile Ala 35 40 45 Glu Ala Asp Gly Asp Ile Val Ala Arg Ala Leu Trp TrpGly Gln Ala 50 55 60 Ala Ser Glu His Pro Val Ala Leu Asp Cys Leu His ValAla Pro Ser 65 70 75 80 Val Pro Asp Arg Ala Gly Leu Ala Ala Asp Leu LeuArg Ala Gly His 85 90 95 Thr Ala Phe Gly Gln Ala Gly Ala Ala Gln Pro ProLeu Tyr Asn Leu 100 105 110 Thr Leu Ser Asn Gly Trp Arg Asp Asp Pro AlaVal Ser Ala Ala Leu 115 120 125 Ala Trp Arg Arg Glu Ala Ala Leu Ala AlaGly Leu Thr His Glu Val 130 135 140 Glu Arg Leu Arg Leu Glu Trp Thr ProGly Thr Gly Val Pro Ala Glu 145 150 155 160 Ser Gly Arg Leu Thr Phe GluGlu Ala Pro Asp Glu Glu Phe Leu Asp 165 170 175 Ala Phe Arg Arg Val AlaGlu Gly Ser Leu Asp Ala Glu Thr Arg Gln 180 185 190 His Leu Ala Ala HisGly Pro Ser Ala Thr Ala Arg Lys Glu Leu Asp 195 200 205 Phe Tyr Leu GlyCys Pro Gly Glu Arg Ala Trp Trp Arg Leu Ala Arg 210 215 220 Thr Pro AspGly Ala Val Ala Gly Phe Ala Ile Pro Ser Ala Thr Pro 225 230 235 240 TyrAsn Pro Asn Val Gly Tyr Leu Gly Val Val Pro Glu Leu Arg Gly 245 250 255Arg Gly Tyr Val Asp Val Leu Leu Ala Glu Ile Thr Arg Ser His Ala 260 265270 Glu Arg Gly Ala Glu Leu Ile Thr Ala Thr Thr Asp Thr Gly Asn Ala 275280 285 Pro Met Ala Ala Ala Phe Glu Arg Ala Gly Tyr Arg Asn Thr Gln Ile290 295 300 Arg Met Leu Phe Ser Ala Pro Ala Gly 305 310 20 341 PRTStreptomyces atroolivaceus 20 Met Thr Gln Thr Ala Pro Leu Ser Pro ArgAsp Arg Pro Ala Ala Thr 1 5 10 15 Pro Glu Pro Ala Ser Arg Pro Tyr AlaArg Gly Tyr Gln Ser Pro Ala 20 25 30 Ala Leu Arg Pro Val Leu Arg Ala AlaHis His Thr Arg Leu Ala Thr 35 40 45 Thr Ala Ala Val Ala Val Ala Asp GlnLeu Val Leu Ile Ala Ala Ile 50 55 60 Leu Ala Ala Ser Trp Pro Gly Trp TrpSer Ala Pro Ala Pro Ala Ala 65 70 75 80 Ala Gly Val Leu Met Ala Leu ThrAla Leu Ile Leu Thr Gly Arg Gln 85 90 95 Ile Arg Ala Leu Glu Cys Leu ValHis Glu Ala Ser His Phe Asn Trp 100 105 110 Ser Arg His His Arg Arg AlaAsn Asp Arg Cys Ala Thr Val Leu Ala 115 120 125 Gly Leu Pro Thr Gly IleArg Ile Ala Arg Tyr Arg Ala Glu His Leu 130 135 140 Val His His Ser ArgLeu Gly Thr Glu Asp Asp Pro Asp Leu Gln Arg 145 150 155 160 Tyr Ala GluLeu Asp Ile Thr Gly Leu Asp Arg Thr Ser Arg Val Gly 165 170 175 Tyr AlaArg Ser Leu Leu Val Arg Ile Ala Ala His Gln Arg Gly Trp 180 185 190 TyrArg Ser Thr Ser Thr Asp Leu Pro Ala Leu Leu Leu Pro Val Gly 195 200 205Trp Ala Leu Leu Val Val Ser Leu Pro Ala Ala Leu Leu Trp Gly Ala 210 215220 Ala Ala Ala Ala Val Ala Ala Gly Val Trp Leu Thr Gly Phe Ala Leu 225230 235 240 Val Leu Pro Val Leu Arg Phe Ile Gly Glu Ser Ser Glu His SerTyr 245 250 255 His Asp Ala Asp Thr Val Phe Asp Ser Thr Ile Thr Asn IleGly Pro 260 265 270 Leu Gln Arg Arg Leu Ile His Pro His Ser Asp Gly TyrHis Thr Val 275 280 285 His His Leu Trp Pro Gly Ala Pro His His Gln LeuArg Leu Leu His 290 295 300 Asp Glu Leu Met Ser Ala Asp Pro Gln Gly PheAla Gly Arg Leu Arg 305 310 315 320 Thr Arg Ser Arg Ile Val Ala Asp ProThr Pro Arg Thr Gly Pro Gly 325 330 335 Ser Thr Pro Trp Thr 340 21 433PRT Streptomyces atroolivaceus 21 Met Arg Pro Asp Met Ser Glu Val AlaVal Glu Gln Leu Lys Phe Leu 1 5 10 15 Ala Thr Asp Asp Val Arg Thr ValAla Glu Glu Phe Gly Thr Pro Val 20 25 30 Phe Val Tyr Asp Glu Ala Thr ValArg Ala Ser Ser Lys Thr Met Ser 35 40 45 Thr Leu Pro Asn Ala Phe Gly LeuThr Val Arg Tyr Ser Leu Lys Ala 50 55 60 Cys Pro Gly Gln Ala Ile Ile ArgLeu Phe Asp Arg Glu Gly Phe Ser 65 70 75 80 Phe Asp Ala Ser Ser Val TrpGlu Ala Arg Arg Ala Val Leu Ala Gly 85 90 95 Val Glu Pro Gly Arg Ile SerMet Thr Ala Gln Glu Ala Val Phe Asp 100 105 110 Glu Asn Leu Arg Asp LeuIle Asp Gln Gly Leu Gln Phe Asp Ala Gly 115 120 125 Ser Leu His Gln LeuGlu Glu Tyr Gly Arg Ala Tyr Pro Gly Gly Ala 130 135 140 Val Ala Ile ArgIle Asn Pro Gly Phe Gly Ser Gly Leu Val Gly Arg 145 150 155 160 Leu AsnSer Gly Gly Pro Gln Ser Ser Phe Gly Ile Trp His Glu Gln 165 170 175 LeuPro Glu Val Lys Gln Ile Val Glu Gln Tyr Gly Leu Arg Leu Val 180 185 190Arg Leu Gln Ser His Ile Gly Ser Gly His His Trp Asp Ile Leu Val 195 200205 Asp Ala Val Arg Thr Met Leu Ser Phe Ala Arg Asp Phe Gln Asp Val 210215 220 His Thr Leu Asp Leu Gly Gly Gly Tyr Arg Val Ser Ala Leu Gln Ser225 230 235 240 Asp Pro Val Tyr Asp His Thr Glu Trp Ala Arg Val Val AlaAsp Glu 245 250 255 Leu Lys Gln Phe Ala Glu Glu Thr Gly Arg Arg Leu HisThr Glu Val 260 265 270 Glu Pro Gly Thr Tyr Leu Thr Ala Leu Ala Gly SerLeu Val Thr Lys 275 280 285 Val Ile Asp Lys Ala Asp Thr Gly Ala Glu GlyAsn Ser Phe Leu Lys 290 295 300 Ile Asn Ser Gly Leu Thr Glu Ile Ala ArgPro Ser Tyr Tyr Gly Val 305 310 315 320 Gln His Pro Ile Val Ser Val ProAla Thr Gly Gly Ala Glu Arg Ala 325 330 335 Ala Asp Asp Arg Glu Glu PheAsn Val Val Gly His Cys Cys Ile Ala 340 345 350 Gly Asp Val Leu Thr AlaLys Gln Glu Glu Leu Ala Pro Val Pro Leu 355 360 365 Gly Arg Thr Glu IleGly Asp Tyr Val Val Ile Glu Arg Ala Gly Ser 370 375 380 Tyr Cys Ser SerMet Ser Met Lys Asn Phe Asn Ser Tyr Pro Glu Ala 385 390 395 400 Ala GluVal Leu Arg Arg Thr Asp Gly Ser Phe Asp Leu Ile Arg Ser 405 410 415 ArgGln Ser Ile Glu Gln Val Ile Val Asn Glu Arg Ile Pro Glu Gly 420 425 430Leu 22 794 PRT Streptomyces atroolivaceus 22 Met Ala Asp Thr Arg Thr ThrPro Ala Asp Leu Thr Ala Ala Glu Glu 1 5 10 15 Pro Arg Pro Pro Ala ProGly Pro Pro Arg Pro Ser Pro Ala Met Arg 20 25 30 Ala Leu Thr Arg Leu LeuVal Pro Val Leu Ala Leu Ala Val Ala Leu 35 40 45 Val Gly Val Leu Trp ThrSer Leu Thr Pro Ala Pro Ala Pro Ala Ser 50 55 60 Ala Pro Ala Gly Ser PheSer Ala Ala Arg Ala Tyr Pro His Leu Ala 65 70 75 80 Ala Val Ser Gly GlyPro His Ala Thr Gly Thr Ala Ala His Asp Arg 85 90 95 Ala Arg Asp Glu ValIle Arg Arg Leu Arg Glu Leu Gly Leu Gly Val 100 105 110 Arg Val Glu ProGly Thr Ser Ser Asp Thr Gly Asn Gly Ala Ala Val 115 120 125 Thr Ala TrpThr Gln Asn Ile Ser Ala Thr Val His Gly Thr His Pro 130 135 140 Ser GlyArg Val Leu Ile Val Ala His Tyr Asp Ser Ala Glu Asn Ser 145 150 155 160His Gly Ala Ser Asp Asp Gly Ile Gly Leu Ala Thr Ala Leu Glu Val 165 170175 Ala Arg Ala Leu Lys Thr Gly Pro Ala Pro Arg Asn Asp Val Thr Phe 180185 190 Leu Ile Thr Asp Gly Glu Glu Pro Gly Leu Leu Gly Ala Arg Ala Phe195 200 205 Val Ala Arg Asp Thr Ala Pro Ala Ala Ser Thr Val Val Leu AsnLeu 210 215 220 Glu Ala Arg Gly Thr Ser Gly Arg Ala Val Met Phe Glu ThrGly Thr 225 230 235 240 Gly Asn Ala Ala Val Val Pro Ala Leu Gly Asp ArgVal Pro Val Ala 245 250 255 Thr Ser Leu Ser Asp Glu Val Tyr Arg Met LeuPro Asn Asp Thr Asp 260 265 270 Phe Thr Val Leu Arg Glu Ala Gly Met ThrGly Met Asn Phe Ala Val 275 280 285 Ile Gly Thr Ser Ala Asn Tyr His ThrPro Gln Asp Asp Leu Ala His 290 295 300 Phe Ser Arg Ala Ser Leu Gln AspMet Gly Asp Thr Val Leu Ala Ala 305 310 315 320 Ala Arg Arg Leu Gly GlyAla Asp Leu Ser Gly Thr Ser His Ala Gly 325 330 335 Gly Ala Thr Tyr PheThr Leu Gly Pro Val Leu Val Arg Tyr Pro Met 340 345 350 Gly Leu Val LeuPro Gly Ala Leu Leu Ala Val Ala Ala Leu Gly Ala 355 360 365 Ala Gly TrpTyr Ala Arg Arg Arg Gly Ala Val Arg Ala Arg Lys Ala 370 375 380 Ala LeuSer Ala Ala Ser Leu Pro Leu Ala Leu Leu Gly Ala Ala Ala 385 390 395 400Ala Val Ser Ala Val Trp Pro Val Ala Thr Trp Leu Arg Pro Gln Trp 405 410415 Thr Gly Phe Ala Leu Gly Ser Pro Tyr Thr Ala Gly Pro Leu His Leu 420425 430 Thr Ala Ala Leu Leu Thr Ala Thr Ala Leu Trp Val Trp Ala Val Val435 440 445 Met Arg Arg Arg Val Ser Ala Leu Glu Ala Gly Ala Ala Val ThrGly 450 455 460 Trp Leu Thr Leu Cys Thr Leu Leu Ser Ala Val Trp Leu ProGly Ala 465 470 475 480 Ser Tyr Leu Phe Leu Trp Pro Ala Leu Thr Gly AlaAla Gly Leu Ala 485 490 495 Ala Ala Ala Arg Ile Ala Pro Gly Ser Pro TrpArg Gly Ala Leu Ala 500 505 510 Thr Ala Pro Leu Val Pro Cys Ala Ala LeuLeu Val Pro Val Gly Ala 515 520 525 Leu Leu Phe Asp Thr Val Pro Ile AlaPhe Ala Ala Ala Pro Ala Leu 530 535 540 Leu Leu Ala Leu Ala Leu Ala ThrAla Leu Pro Ala Val Pro Ala Lys 545 550 555 560 Thr Arg Arg Arg Ser LeuVal Thr Gly Thr Gly Leu Gly Thr Ala Ala 565 570 575 Ala Leu Ala Ala LeuGly Ala Ala Leu Leu Ala Gly Ala Pro Thr Ala 580 585 590 Ser Arg Pro AlaGln Val Ser Leu Leu Tyr Ala Leu Asp Ala Asp Thr 595 600 605 Gly Arg AlaArg Trp Ala Ser Ser Gly Ala Gly Asp Ser Ala Trp Leu 610 615 620 Asp HisTyr Val Gly Gly Ala Thr Thr Ser Leu Glu Ser Thr Phe Pro 625 630 635 640Ser Leu Asp Gln Pro Ser Arg Trp His Thr Gly Pro Ala Pro Ala Ala 645 650655 Thr Val Ala Val Pro Glu Val Lys Val Val Ser Thr Thr Arg Ala Gly 660665 670 Asp Val Arg Arg Ile Ile Leu Ser Val Arg Ala Thr Asp Ala Arg Ala675 680 685 Ala Glu Ile Gly Leu Tyr Ala Asp Ala Gly Gly Ala Ala Val ValArg 690 695 700 Ala Glu Val Ala Gly Gln Ser Val Pro Gly Gly Thr Asn ArgPro Phe 705 710 715 720 Ala Ser Asp Gly Trp Arg Trp Gly Leu Leu Leu AlaAla Pro Pro Thr 725 730 735 Gly Pro Gly Thr Arg Val Glu Leu Thr Leu ArgGly Thr Gln Pro Leu 740 745 750 Thr Leu Arg Ala Val Thr Arg Thr Pro GlyLeu Pro Gly Gly Thr Leu 755 760 765 Ser Val Pro Arg Pro Ala Asp Val ThrTrp Ser Ala Asp Asp Ser Gly 770 775 780 Leu Thr Leu Val Ser Arg Arg TyrVal Leu 785 790 23 432 PRT Streptomyces atroolivaceus 23 Met Ala Pro ArgThr Pro Arg Phe Ala Tyr Gly Tyr Gly Arg Leu Trp 1 5 10 15 Ala Ala ThrAla Ala Ser Gly Phe Gly Asp Gly Thr Arg Thr Ala Ala 20 25 30 Leu Ala ValTyr Ala Ala Thr Leu Thr His Asp Pro Phe Ala Val Ala 35 40 45 Leu Val AlaMet Ala Gly Lys Leu Pro Trp Thr Cys Val Gly Pro Phe 50 55 60 Ala Gly AlaLeu Val Asp Arg Leu Asp Arg Trp Arg Thr Leu Trp Ile 65 70 75 80 Cys AspVal Ala Arg Val Leu Val Thr Gly Ala Leu Val Leu Leu Ile 85 90 95 Leu ThrAsp Arg Ala Gly Ile Thr Ala Leu Ala Val Val Ser Phe Leu 100 105 110 LeuThr Ser Ile Thr Thr Leu Ala Glu Asn Leu Ser Gln Ala Val Val 115 120 125Pro Asp Val Thr Gly Gly Gly Ser Leu Asp Ser Ala Asn Ser Arg Leu 130 135140 Met Gly Gly Gln Leu Val Ser Ser Glu Phe Leu Gly Ala Pro Leu Gly 145150 155 160 Thr Ala Leu Phe Ala Leu Gly Arg Pro Val Pro Phe Ala Val AspThr 165 170 175 Leu Ser Phe Ala Val Ser Ala Val Leu Val Phe Ser Ile ArgLeu Ser 180 185 190 Gly Ala Arg Pro Ala Pro Thr Ala Ala Gly Arg Leu ThrAla Arg Ala 195 200 205 Val Val Ala Glu Thr Ala Asp Gly Ile Arg Trp LeuTrp Arg His Arg 210 215 220 Val Leu Arg Thr Val Cys Val Leu Val Gly AlaLeu Asn Phe Ala Val 225 230 235 240 Val Ala Val Leu Ser Ile Ala Val LeuTyr Ala Leu Asn Val Leu His 245 250 255 Ile Ser Gln Arg Ala Tyr Gly LeuLeu Leu Ala Val Leu Ala Val Gly 260 265 270 Gly Leu Ala Gly Val Leu LeuAla Pro Leu Val Thr Arg Ala Ile Gly 275 280 285 Arg Ala Arg Thr Leu GlnLeu Ala Phe Ala Leu Cys Pro Leu Pro Phe 290 295 300 Leu Ile Ala Gly LeuThr Ser Leu Pro Leu Leu Ala Ala Gly Ala Leu 305 310 315 320 Ala Phe ValGly Ala Ala Ile Ser Leu Ala Thr Val Val Thr Thr Ser 325 330 335 Leu ArgGln Ser Leu Val Pro Gly Glu Leu Phe Gly Arg Val Asn Gly 340 345 350 GlyTyr Arg Phe Val Val Asn Gly Val Ser Pro Leu Gly Gly Val Ala 355 360 365Gly Gly Ala Leu Ala Glu Ala Ala Gly Leu Arg Ala Pro Phe Leu Phe 370 375380 Ala Ala Ala Leu Leu Ala Ala Ala Thr Val Thr Ala Leu Leu Arg Leu 385390 395 400 Ser Pro Ala Ala Leu Asp Ala Ala Gly Asp Asp Arg Pro Ala ProArg 405 410 415 Thr Gly Gln Ala Val Ala Ala Glu Arg Thr Glu Ala Pro GluArg Thr 420 425 430 24 1134 PRT Streptomyces atroolivaceus 24 Met ThrGlu Thr Leu Pro Ala Asp Ala Ser Ala Leu Gln Arg Ala Val 1 5 10 15 AlaGlu Leu Ser Pro Glu Arg Arg Ala Leu Leu Ala Arg His Leu Ala 20 25 30 ArgGly Arg Ala Gly Ser Gly Ile Ser Val Thr Ala Leu Pro Arg Thr 35 40 45 GlyGlu Ala Gln Arg Tyr Val Ala Ser Pro Gly Gln Gln Gln Leu Tyr 50 55 60 TyrGln Tyr Gln Leu Asp Pro Ser Ala Val Asn Tyr Leu Leu Pro Ile 65 70 75 80Ala Val Arg Leu Arg Gly Asp Leu Asp Thr Gln Ala Leu Glu Arg Ala 85 90 95Leu Ala Ala Val Val Ala Arg His Glu Ala Leu Arg Thr Arg Phe Ala 100 105110 Thr Asp Pro Glu His Gly Val Val Gln Val Val Gly Thr Pro Asp Glu 115120 125 Val Arg Val Ala Leu Gly Arg Gly Arg Ala Arg Ala Asp Glu Val Pro130 135 140 Ala Leu Leu Asp Arg Ile Ala Arg Arg Pro Phe Asp Leu Ser AlaAla 145 150 155 160 Pro Leu Leu Arg Ala Lys Leu Trp Gln Leu Thr Asp AspPro Glu Ala 165 170 175 Pro Asp Gly Arg Ser Trp Leu Leu Thr Leu Cys ValHis His Ile Val 180 185 190 Val Asp Gly Trp Ser Leu Gly Val Leu Val AspGlu Leu Thr Glu Gly 195 200 205 Tyr Ala Ala Gln Leu Thr Gly Arg Pro AlaAsn Leu Pro Glu Leu Ala 210 215 220 Val Gln Tyr Pro Asp Phe Ala Ala AlaHis His Ala Trp Leu Ala Gly 225 230 235 240 Asp Glu Ala Ala Arg Gln IleAla His Trp Arg Glu Gln Leu Asp Gly 245 250 255 Val Leu Pro Leu Glu LeuAla Gly Asp Arg Pro Arg Pro Ala Arg Pro 260 265 270 Thr Phe Ser Gly AlaAla Val Arg Ile Asp Leu Asp Ala Gln Leu Val 275 280 285 Glu Arg Ala AspArg Leu Gly Arg Thr Glu Lys Ala Thr Pro Phe Met 290 295 300 Val Leu LeuAla Ala Tyr Ala Val Val Leu His Arg Trp Ser Ala Ala 305 310 315 320 ArgAsp Leu Val Ile Gly Thr Ala Val Ala Gly Arg Thr Arg Pro Glu 325 330 335Ser Glu Pro Leu Ile Gly Phe Phe Val Asn Thr Leu Pro Leu Arg Val 340 345350 Pro Val Asp Gly Ala Ala Ser Phe Arg Glu Leu Leu Arg Ala Thr Arg 355360 365 Asp Ser Ser Leu Asp Gly Phe Ala His Gln Asp Val Pro Phe Pro Arg370 375 380 Ile Val Arg Glu Val Gly Gly Glu Arg Thr Ser Gly Arg Ser AlaLeu 385 390 395 400 Val Arg Ala Leu Phe Ala Leu Arg Asn Val Pro Leu ArgHis Leu Glu 405 410 415 Leu Ala Gly Val Arg Ala Glu Thr Leu Asn Leu ProLys Thr Gly Ser 420 425 430 Asp Ile Asp Leu Ser Leu Glu Phe Ser Pro AlaGlu His Gly Gly Gly 435 440 445 Leu Thr Gly Trp Leu Ser Tyr Ser Phe AspLeu Phe Asp Gln Asp Thr 450 455 460 Ala Glu Arg Ile Ala Arg Ala Val HisGln Val Leu Thr Ala Ala Ile 465 470 475 480 Ala Ser Asp Gly Asp Thr AlaVal Arg Asp Leu Pro Val Leu Thr Glu 485 490 495 Glu Glu His Thr Thr LeuAla Leu His Trp Ser Gly Arg Asp Ala Pro 500 505 510 Ala Pro Gly Gly AspGly Leu Leu Asp Ala Phe His Ala His Ala Ala 515 520 525 Ala Thr Pro AspAla Thr Ala Val Val Ala Asp Val Pro Pro Pro Ala 530 535 540 Thr Gly HisArg Leu Thr Tyr Arg Glu Leu Asp Glu Arg Ala Gly Arg 545 550 555 560 LeuAla His Leu Leu Arg Ala Arg Gly Ile Gly Pro Glu Asp Arg Val 565 570 575Ala Leu Leu Leu Pro Arg Gly Leu Asp Leu Leu Val Cys Leu Leu Ala 580 585590 Val Leu Lys Ala Gly Ala Val Leu Val Pro Leu Glu Ser Glu His Pro 595600 605 Ala Gly Arg Leu Ala Gly Ile Val Ala Asp Ala Thr Pro Cys Leu Thr610 615 620 Leu Thr Asp Thr Arg Leu Ala Ala Arg Leu Pro Glu Gly Thr ProVal 625 630 635 640 Leu Thr Val Glu Asp Leu Pro Ala Leu Thr Ala His HisPro Ala Thr 645 650 655 Gly Pro Asp Val Pro Glu Leu Pro Ala Ser Gly AlaTyr Val Leu Tyr 660 665 670 Thr Ser Gly Ser Thr Gly Ala Pro Lys Gly ValLeu Ile Thr Arg Ala 675 680 685 Gly Leu Ala Ser Arg Leu Ala Gly Met CysAla Asp Leu Gly Leu Gly 690 695 700 Ala Asp Glu Arg Val Leu His Lys ThrPro Leu Ser Ala Asp Thr Ser 705 710 715 720 Met Trp Glu Leu Leu Val AlaLeu Arg Ser Gly Gly Arg Val Val Leu 725 730 735 Ala Ala Pro Gly Leu GlyThr Asp Val Asp Tyr Leu Tyr Ser Val Leu 740 745 750 Thr Arg His Ala ValThr Thr Cys Phe Phe Val Pro Ser Ala Leu Arg 755 760 765 Pro Ala Ile GlyLeu Gly Gly Leu Pro Arg Ala Ala Ala Leu Arg Leu 770 775 780 Val Ile SerAla Gly Glu Glu Leu Pro Ala Asp Leu Ala Asp Gln Leu 785 790 795 800 LeuLeu Gln Val Pro His Ile Arg Leu Val Asn Ser Tyr Gly Pro Thr 805 810 815Glu Thr Thr Ile Asn Ile Ala Glu His Thr Val Thr Ala Pro Ala Ala 820 825830 Ser Pro Val Pro Ile Gly Arg Pro Val Pro Gly Gly Asp Leu Tyr Val 835840 845 Leu Asp Gly Ala Gly Arg Val Gln Pro Val Gly Val Pro Gly Glu Leu850 855 860 His Ala Gly Gly Val Gln Val Ala Arg Gly Tyr Leu Gly Arg ProAla 865 870 875 880 Gln Thr Ala Glu Val Tyr Val Pro His Pro Phe Val ProGly Ala Arg 885 890 895 Val Tyr Arg Thr Gly Asp Arg Ala Arg Trp Arg ProAsp Gly Thr Leu 900 905 910 Glu Phe Leu Gly Arg Thr Asp His Gln Val LysIle Arg Gly Phe Arg 915 920 925 Val Glu Pro Ala Glu Val Glu Thr Ala LeuArg Ala His Pro Gln Val 930 935 940 Ala Asp Ala Leu Val Leu Ala Asp AlaAla Pro Asp Gly Gly Ala Arg 945 950 955 960 Leu Ile Gly Tyr Val Thr ValAla Pro Gly Gln Ser Ala Pro Asp Thr 965 970 975 Pro Ala Leu Arg Thr HisLeu Arg Ala Ala Leu Pro Ser Pro Met Val 980 985 990 Pro Asp Val Phe ValAla Leu Asp Ala Phe Pro Leu Thr Pro Tyr Gly 995 1000 1005 Lys Ile AspArg Ala Ser Leu Pro Arg Pro Gly Arg Gly Pro Ala 1010 1015 1020 Lys GluArg Arg Ala Pro Ala Asp Asp Leu Gln His Val Leu Ala 1025 1030 1035 GlyVal Trp Glu Gln Thr Leu Glu Ile Thr Asp Val Ser Val Asp 1040 1045 1050Asp Asp Phe Phe Glu Arg Gly Gly His Ser Val Leu Ala Thr Ile 1055 10601065 Val Val Ser Ser Ile Arg Glu Leu Phe Arg Leu Glu Leu Pro Leu 10701075 1080 His Phe Phe Val Glu Ala Pro Thr Val Ala Thr Leu Ala Thr Leu1085 1090 1095 Met Arg Ala Glu Gly Ala Glu Ala Gly Ile Asp Val Asp ArgVal 1100 1105 1110 Ala Ala Leu Val Arg Gln Val Gln Ser Met Ser His AsnGlu Val 1115 1120 1125 Gln Gln Lys Leu Glu Gly 1130 25 276 PRTStreptomyces atroolivaceus 25 Met Arg Ser Val Arg Thr Ala Ala Gly AlaLeu Ser Pro Ala Ala His 1 5 10 15 Arg Lys Gly Leu Asp Leu Val Ser GluThr Thr Thr Ala Ala Ala Gln 20 25 30 Thr Arg Thr Val Thr Ala Ala Val ThrGly Leu Ser Gly Trp Gly Gly 35 40 45 His Leu Leu Thr Ala Pro Glu Val ArgArg Leu Thr Gly Thr Gly Pro 50 55 60 Gln Asp Trp Ala Arg Phe Ala Arg HisTrp Asp Asp Leu Thr Leu Asp 65 70 75 80 Thr Phe Met Lys Asp Gly Gly ThrTyr Arg Tyr Arg Arg Tyr Gly His 85 90 95 Phe Arg Leu Asp Val Arg Thr GlyGlu Leu Thr Ala Gln Pro His Ala 100 105 110 Pro Tyr Phe Gln Asp Ser AlaLeu Asn Pro Leu Asn Gly Gly Val Gln 115 120 125 Arg Ser Phe Asp Pro LeuThr Glu Ala Phe Leu Ala Asp Pro Leu Thr 130 135 140 Ala Ala Val Val HisLeu Leu Gly Asp Val Phe Ser Arg Ala Glu Gly 145 150 155 160 Val Arg LeuTrp Asp Val Lys Leu His Pro Phe Arg Ile Val Thr Ser 165 170 175 Pro AspGln Thr Gly Arg Pro Ala Pro Gln Gly Arg His Arg Asp Gly 180 185 190 GlyAla Tyr Val Thr Ser Leu Leu Val Asn Arg Thr Asn Val Ser Gly 195 200 205Gly Glu Ser Ser Leu Tyr Asp Asp Asp Gly Asn Gln Leu Phe Ala Val 210 215220 Thr Leu Gly Glu Pro Gly Asp Gln Leu Leu Val Asp Asp Arg Lys Val 225230 235 240 Leu His Asp Val Thr Pro Leu Ala Pro Val Asp Pro Ala Arg ProSer 245 250 255 His Arg Asp Val Leu Ile Val Asp Phe Asp Pro Val Ala AlaAsp Ala 260 265 270 Glu Gly Gln Arg 275 26 549 PRT Streptomycesatroolivaceus 26 Met Thr Ala Asp Asp Ser Leu Thr Val Val Thr Ser Phe AlaGln Gln 1 5 10 15 Ser Leu Trp Leu Gln His Gln Val Asp Pro Asp Arg AlaAla Tyr Asn 20 25 30 Ile Thr Ala Ala Val Arg Leu Arg Gly Pro Leu Asp SerAla Ala Leu 35 40 45 Glu Arg Ala Leu Asn Ala Val Val Ala Arg His Glu ValLeu Arg Thr 50 55 60 Asp Phe Ala Leu Asp Gly Glu Asp Pro Val Gln Val ValArg Pro Glu 65 70 75 80 Arg Glu Leu Arg Val Pro Val Leu Asp Ala Gly ProAsp Asp Phe Leu 85 90 95 Arg Tyr Ala Gly Lys Val Leu Phe Ala Pro Phe AspLeu Arg Gly Gly 100 105 110 Pro Leu Leu Arg Leu Glu Leu Leu Arg Leu GlyPro Glu His His Ile 115 120 125 Ala Leu Leu Ala Leu His His Ile Val ThrAsp Gly Ala Ser Ser Ala 130 135 140 Ile Leu Leu Gln Glu Leu Ser His AlaTyr Ala Ala Glu Leu Ala Gly 145 150 155 160 Val Glu Pro Leu Trp Glu GluLeu Pro Ile Gln Tyr Ala Asp Phe Ala 165 170 175 Val Trp Gln Arg Asp AlaLeu Ser Gly Ala Ala Leu Asp Glu Leu Thr 180 185 190 Gly His Trp Thr ArgThr Leu Ala Gly Ala Glu Pro Leu Ala Leu Pro 195 200 205 Ala Asp Arg GluArg Asp Ala Asp Ala Ser Gly His Ala Ala Val His 210 215 220 His Phe ThrLeu Pro Ala Ala Leu Thr Ala Arg Leu Glu Asp Leu Ala 225 230 235 240 ArgAla Glu Arg Ala Thr Leu Phe Met Val Leu Leu Ala Gly Leu Asp 245 250 255Ala Leu Leu Ala Arg Tyr Cys Gly Gln Glu Asp Ile Thr Val Val Ser 260 265270 Pro Met Ala Gly Arg Gly Arg Pro Glu Leu Glu Gly Leu Ile Gly Tyr 275280 285 Phe Val Asn Pro Leu Leu Leu Arg Thr Gln Val Ser Gly Thr Ala Gly290 295 300 Phe Ala Ala Leu Val Gly Arg Thr Arg Asp Thr Cys Leu Asp AlaPhe 305 310 315 320 Asp Arg Glu Glu Leu Pro Phe Glu Gln Ala Val Glu ValLeu Arg Gly 325 330 335 His Gly Ala Pro Gly Ala Glu Ser Leu Arg Ser GlnVal Met Leu Val 340 345 350 Leu Gln Asn Thr Arg Pro Asn His Trp His SerAla Gly Ile Asp Phe 355 360 365 Glu Leu Ile Pro Ala Asp Thr Ser Thr AlaLys Ser Asp Leu Val Leu 370 375 380 Glu Val Arg Pro Gly Ala Asp Gly GlyCys Thr Ala Ala Leu Glu Tyr 385 390 395 400 Arg Thr Gly Leu Tyr Asp AlaAla Thr Ala Asp Arg Val Ala Arg His 405 410 415 Phe Ala Ala Ile Leu GluAla Ala Cys Glu Arg Pro Asp Glu Pro Leu 420 425 430 Ser Gln Leu Leu AlaThr Thr Val Pro Asp Gly Glu Ile Pro Ala Val 435 440 445 Arg Ser Gly AlaAla His Gly Glu Ala Pro Ala Ala Thr Glu Tyr Val 450 455 460 Ala Pro ArgThr Pro Leu Glu Glu Glu Val Ala Gly Ile Trp Ser Ala 465 470 475 480 LeuLeu Gly Thr Glu Arg Val Gly Val His Asp Asn Phe Phe Asp Leu 485 490 495Gly Gly Gln Ser Leu Leu Ala Val Arg Leu Ala Ala Arg Leu Arg Asp 500 505510 Glu Phe Gly Val Glu Met Thr Val Arg Asp Leu Tyr Ala Asp Phe Thr 515520 525 Leu Glu Ala Val Thr Trp Ala Val Leu Gln Arg Met Thr Ala Asp Gly530 535 540 Glu Gln Ala Pro Ala 545 27 235 PRT Streptomycesatroolivaceus 27 Met Leu Ser Ala Ala Val Leu Cys Phe Ala Pro Ala Gly AlaGly Ala 1 5 10 15 Ser Phe Phe His Pro Trp Leu Gly Gln Arg Glu Gly LeuThr Val Leu 20 25 30 Pro Val Glu Leu Pro Gly Arg Glu Lys Arg Phe Ala GluPro Glu Cys 35 40 45 Thr Glu Leu Gly Ala Leu Met Asp Ala Val Leu Pro AlaLeu Arg Ser 50 55 60 Ala Thr Ala Gly Phe Asp Lys Val Ala Val Leu Gly HisSer Phe Gly 65 70 75 80 Ala Leu Leu Ala Tyr Glu Ala Thr Arg Ala Leu AlaAla Glu Arg Asp 85 90 95 Gly Leu Thr Leu Val Ala Ser Gly Ala Thr Trp ProGly Thr Leu Arg 100 105 110 Arg Gly Arg Ile Thr Gly Leu Ala Asp Asp GluPhe Ile Ala Gly Ile 115 120 125 His Thr Ile Ala Gly Tyr Arg His Pro AlaLeu Asp Glu Pro Glu Leu 130 135 140 Arg Asp Leu Val Leu Pro Pro Leu ArgThr Asp Val Ala Met His Glu 145 150 155 160 Asn Tyr Val Tyr Val Glu ArgGlu Pro Leu Asp Val Pro Val Leu Ala 165 170 175 Val Arg Gly Asp Ala AspArg Leu Val Ser Ala Asp Asp Leu Thr Glu 180 185 190 Trp Glu Arg Val ThrThr Gly Thr Val Thr Val Ala Glu Leu Thr Gly 195 200 205 Ala His Met TyrLeu Val Glu Glu Trp Pro Ala Leu Leu Asp Leu Leu 210 215 220 Ala Gly HisLeu Leu Gly Ala Gly Thr Pro Ser 225 230 235 28 276 PRT Streptomycesatroolivaceus 28 Met Ser Gly Arg Leu Thr Gly Arg Thr Ala Ile Val Thr GlyAla Gly 1 5 10 15 Arg Gly Val Gly Arg Ala Cys Ala Thr Ala Phe Ala AlaGln Gly Ala 20 25 30 Asp Leu Val Leu Val Asp Ile Ala Ala Asp Leu Pro HisVal Pro Tyr 35 40 45 Pro Ala Ala Thr Pro Ser Gln Leu Asp His Thr Ala ArgLeu Cys Arg 50 55 60 Glu Gln Gly Ala Ala Val Leu Thr Ala Arg Ala Asp ValArg Asp Ala 65 70 75 80 Ala Ala Cys Glu Arg Val Val Ala Asp Ala Val AspArg Phe Gly Ser 85 90 95 Leu Asp Val Leu Val Asn Asn Ala Gly Ile Ala GlyPro Ser Gly Arg 100 105 110 Ile Val His Glu Val Thr Glu Asp Glu Trp AlaVal Met Ile Asp Val 115 120 125 Asn Leu Asn Gly Ala Trp Arg Met Leu LysAla Ala Gly Ala Ser Met 130 135 140 Val Ala Ala Arg Ser Gly Ser Ile ValAsn Ile Ala Ser Thr Ala Gly 145 150 155 160 Leu Val Gly Tyr Arg Asn PheAla Gly Tyr Val Ala Ser Lys His Gly 165 170 175 Leu Val Gly Leu Thr LysAla Ala Ala Leu Asp Tyr Ala Pro Tyr Arg 180 185 190 Val Arg Val Asn AlaVal Cys Pro Gly Ser Val Arg Asp Gly Glu Ala 195 200 205 Trp Glu Gly ArgMet Leu Val Glu Ile Gly Arg Ser Ile Gly Ile Glu 210 215 220 Pro Ala AspHis Glu Ala Glu Phe Ile Thr Gln Met Pro Met Asn Thr 225 230 235 240 LeuVal Glu Ala Asp Asp Val Ala Gly Ala Ala Leu Trp Leu Ala Ser 245 250 255Glu Glu Ser Arg His Ser Thr Gly Gly Val Ile Thr Val Asp Ala Gly 260 265270 Tyr Ser Ala Arg 275 29 920 PRT Streptomyces atroolivaceus 29 Met SerSer Asn Ser Pro Ser Val Ala Ala Arg His Ala Ala Val Tyr 1 5 10 15 AlaVal Ala Pro Gly Tyr Ser Ala Glu Asp Cys Arg Ala Ala Leu Ala 20 25 30 GlyAla Pro Trp Pro Gly Pro Gly Arg Leu Arg Leu Trp Ala Glu Pro 35 40 45 ValPro Ala Pro Ser Asp Ser Pro Ala Ala Leu Arg Arg Arg Thr Arg 50 55 60 GluLeu His Gly Arg His Gly Gly Glu Gly Val Arg Leu Thr Leu Leu 65 70 75 80Glu Tyr Ala Asp Gly Val Arg Asp Leu Val Ala Val Thr Gly Pro Gly 85 90 95Asp Asp Gly Ala Asp Arg Leu Asp Arg Ala Ala Leu Ala Ala Leu Gly 100 105110 Pro Ala Arg Pro Leu Ala Gly Ser Gly Ser Pro Ser Asp Pro Gly Cys 115120 125 Arg Ala Val Pro Val Pro Gly Leu Gly Ala Gly Gly Ala Met Ala Thr130 135 140 Ala Trp Asp Leu Thr Ala Ala Leu Leu Leu Ile Ala Ala Arg ThrArg 145 150 155 160 Ala Gln Asp Glu Pro Val Leu Asp Val Ala Val His ThrThr Asp Gly 165 170 175 Val Arg Arg Val Ser Leu Arg Val Ala Ala Leu AspGlu Asp Gln Pro 180 185 190 Val Ser Ala Tyr Arg Ala Trp Val Arg Glu GlnAla Glu Gln Thr Ser 195 200 205 Ala Gln Asp Gly Gly Gly Gly Pro Arg LeuAla Leu Asp Val Asp Leu 210 215 220 Arg Pro Arg Ala Val Pro Ala Val ArgArg Leu Pro Ala Leu His Ala 225 230 235 240 Arg His Pro Val Thr Val ArgVal Thr Ala Ser Asp Ala Asp Gly Thr 245 250 255 Pro Gln Val His Cys TrpThr Leu Pro Gly Thr Pro Ala Asp Gln Leu 260 265 270 Pro Thr Asp Phe GluGln Gln Leu Ala Val Val Ala His Gln Leu Leu 275 280 285 Cys Gly Asp GlnAsn Leu Pro Leu Ala Asp Val Ala Leu His Glu Pro 290 295 300 Ala Glu ArgAla Arg Ile Leu Asp Leu Gly Arg Thr Pro Arg Ala Ala 305 310 315 320 AspSer Gly Pro Arg Arg Ile Asp Gln Leu Val Arg Asp Arg Ala Glu 325 330 335Arg Thr Pro Asp Ala Val Ala Leu Arg Asp Pro Gln Gly Glu His Thr 340 345350 Trp Thr Tyr Gly Glu Leu Val Asp Arg Ser Asp Arg Phe Ala Ala Ala 355360 365 Leu Arg Gly Leu Gly Val Arg Pro Gly Asp Arg Val Gly Val Cys Leu370 375 380 Asp Arg Ser Ala Gln Leu Val Ser Val Leu Leu Ala Val Met ThrAla 385 390 395 400 Gly Ala Ala Tyr Val Pro Leu Asp Pro Thr Tyr Pro AlaAsp Arg Leu 405 410 415 Ala Tyr Thr Ala Asp Asp Ala Gly Leu Ser Leu ValVal Val Glu Asp 420 425 430 Gly Gly Lys Asp Asp Gly Asn Ala Phe Ala AspHis Ala Thr Val Thr 435 440 445 Leu Pro Arg Leu Arg Glu Leu Ala Ala GlyGln Gly Ala Trp Glu Glu 450 455 460 Pro Gly Thr Val Gly Pro Asp Asp ProAla Tyr Ile Ile Tyr Thr Ser 465 470 475 480 Gly Ser Thr Gly Arg Pro LysGly Val Val Val Pro His Arg Asn Val 485 490 495 Gly Arg Leu Leu Asp AlaThr Ala Asp Asp Phe Arg Leu Gly Pro Gln 500 505 510 Asp Val Trp Thr TrpPhe His Ser Ala Ala Phe Asp Phe Ser Val Trp 515 520 525 Glu Ile Trp GlyAla Leu Gly Thr Gly Gly Arg Leu Val Val Val Pro 530 535 540 Tyr Trp ThrCys Arg Ser Pro Glu Asp Phe Arg Ala Leu Leu Leu Asp 545 550 555 560 GluArg Val Thr Val Leu Asn Gln Thr Pro Ser Ala Phe Ser Arg Leu 565 570 575Leu Pro Leu Glu Arg Ala Ala Pro Thr Pro Leu Ala Leu Arg Leu Val 580 585590 Val Phe Gly Gly Glu Pro Leu Asp Ala Arg Ala Leu Leu Pro Trp Phe 595600 605 Asp Thr His Pro Glu Ser Ala Cys Arg Met Val Asn Met Phe Gly Ile610 615 620 Thr Glu Thr Thr Val His Val Thr Ala Gln Thr Val Thr Arg AlaAsp 625 630 635 640 Ala Leu Ala Ser Ser Gln Ser Val Gly Arg Ala Leu ProGly Trp Ser 645 650 655 Val Arg Val Leu Asp Thr Arg Gly Arg Leu Val GlnPro Gly Cys Val 660 665 670 Gly Glu Ile Ala Val Gly Gly Asp Gly Leu AlaLeu Glu Tyr Leu Gly 675 680 685 Arg Pro Glu Leu Thr Ala Glu Arg Phe ValPro Asp Pro Asp Gly Glu 690 695 700 Gly Arg Leu Tyr Leu Ser Gly Asp LysGly Arg Gln Leu Pro Asp Gly 705 710 715 720 Arg Leu Glu His Leu Gly ArgLeu Asp Ser Gln Val Lys Leu Arg Gly 725 730 735 His Arg Ile Glu Leu AspGlu Ile Arg Ser Val Leu Leu Thr His Pro 740 745 750 Ser Val Arg Ala AlaAla Val Val Leu Thr Arg Pro Thr Asp Ala Asn 755 760 765 Gly Glu Asp AlaThr Leu Asp Ala Tyr Ala Val Leu Asp Gly Ala Asp 770 775 780 Ala Arg GluVal Arg Arg His Ala Ala Arg Leu Leu Pro Glu Tyr Met 785 790 795 800 ValPro Ala Thr Val Thr Pro Leu Ala Glu Leu Pro Leu Thr Val Asn 805 810 815Gly Lys Val Asp Val Ala Ala Leu Pro Ala Pro Arg Ala Ala Gly Gly 820 825830 Glu Leu Pro Cys Val Ala Asp Glu Ser Ala Gln Gly Thr Leu Ala Ala 835840 845 Val Leu Ala Ala Trp His Thr Ala Phe Gly Glu Gln Val Ala Ala Gly850 855 860 Asp Asp Phe Phe Glu Leu Gly Gly Asn Ser Leu Arg Ala Leu ArgVal 865 870 875 880 Val His Leu Leu Arg Asp Ala Gly Leu Arg Val Asp ValArg Asp Val 885 890 895 Tyr Arg Leu Arg Thr Pro Ser Ala Leu Ala Glu AlaAla Ser Thr Ala 900 905 910 Pro Ala Asp Ala Ala Arg Thr Ala 915 920 30195 PRT Streptomyces atroolivaceus 30 Met Ser Lys Val Ala Val Leu ThrAsn Asp Leu Gln Tyr Asp Leu Val 1 5 10 15 Asn Lys Asn Gln Glu Arg IleAsp Ala Val Ala Glu Ala Thr Pro Ser 20 25 30 Phe Ser Gly Phe Leu Lys AspMet Arg Gln Arg Gly His His Val Leu 35 40 45 His Leu Gln Leu Ile Asn LysAla Asp Asp Pro Val Ala Glu Arg Tyr 50 55 60 Glu Gly Gly Arg Leu Pro AlaGln Lys Gly Thr Gln Gly Ala Glu Ile 65 70 75 80 Leu Pro Val Phe Leu AspAsp Thr Asp Ile Leu Val Glu Lys Asn Lys 85 90 95 Asp Ser Gly Phe Tyr GluThr Asp Leu His Glu Arg Leu Gln Ala Leu 100 105 110 Gly Val Glu Thr ValIle Val Thr Gly Met Gln Gly Gln Ile Cys Val 115 120 125 Gln Thr Thr AlaAla Asp Ala Phe Phe Arg Gly Tyr Asn Val Trp Val 130 135 140 Pro Ser AspCys Val Val Ser Ala Gln Glu His Asp Lys Val Arg Ala 145 150 155 160 LeuAsp Trp Leu Asp Gly Tyr Cys Ala Thr Val Ala Pro Ser Asp Glu 165 170 175Ile Val Arg Val Leu Asp Ala Asp Gly Asp Leu Pro Arg Lys Val Phe 180 185190 Arg Thr Pro 195 31 343 PRT Streptomyces atroolivaceus 31 Met Pro ThrAla Thr Thr Ser Phe Leu Asp Gln Lys Ile Val Leu Asp 1 5 10 15 His GlyThr Gly Ala Arg Leu Ser Arg Glu Leu Val Glu Phe Val Val 20 25 30 Glu ValLeu Gly Asp Val Tyr Leu Gly Glu Met Glu Asp Ser Ala Ile 35 40 45 Leu ProPro Leu Asp Gly Ala Leu Ala Met Thr Thr Asp Ser Phe Val 50 55 60 Val AspPro Pro Phe Phe Gly Asn Gly Asp Ile Gly Lys Ile Ala Val 65 70 75 80 CysGly Thr Val Asn Asp Leu Ala Val Ala Gly Ala Glu Pro Arg Tyr 85 90 95 LeuThr Leu Ala Met Ile Leu Glu Ser Gly Phe Pro Leu Ser Gly Leu 100 105 110His Arg Ala Leu Glu Ser Val Arg Asp Thr Ala Arg Glu Ala Gly Ile 115 120125 Lys Ile Val Ala Gly Asp Thr Lys Val Val Gly Glu Gly Glu Ala Asp 130135 140 Gly Ile Phe Leu Asn Thr Thr Gly Val Gly Val Phe Thr Ser Pro Pro145 150 155 160 Lys Arg Met Lys Asp Val Ala Val Gly Asp Lys Ile Leu LeuSer Gly 165 170 175 Pro Ile Gly Asp His Thr Ile His Leu Leu Ser Val ArgGlu Gly Leu 180 185 190 Gly Phe Glu Lys Asn Val Leu Ser Asp Cys Ala ProLeu Asn Asn Met 195 200 205 Ile Asp Thr Val Arg Gly Ala Val Pro Glu SerAla Val Arg Ser Met 210 215 220 Arg Asp Val Thr Arg Gly Gly Leu Asn AlaVal Leu His Glu Tyr Ala 225 230 235 240 Ser Ala Leu Gly His Ala Val ArgVal Glu Glu Glu Ala Leu Pro Ile 245 250 255 Ser Phe Glu Thr Ala Met AlaAla Asp Met Leu Gly Ile Asn Pro Ile 260 265 270 Asn Cys Ala Asn Glu GlyCys Leu Ala Leu Phe Val Ala Pro Glu His 275 280 285 Ala Asp Glu Val LeuArg Val Leu Arg Ser His Pro Tyr Gly Lys Asp 290 295 300 Ala Val Ile ValGly Glu Val Thr Ala Arg Pro Glu Pro Val Val Glu 305 310 315 320 Met AlaGly Lys Asp Gly Arg Ile Gln Glu Ile Glu Glu Leu Gln Gly 325 330 335 AlaGlu Leu Pro Arg Leu Cys 340 32 791 PRT Streptomyces atroolivaceus 32 MetAla Glu Thr Glu Gln Trp Leu Ile Arg Val Thr Gly Val Val Gln 1 5 10 15Gly Val Gly Tyr Arg Pro Phe Val His Thr Leu Ala Thr Gly Leu Gly 20 25 30Leu Arg Gly Trp Val Leu Asn Asp Thr Leu Gly Val Gln Thr Glu Val 35 40 45Ser Gly Gly Ala Gly Glu Leu Ala Glu Phe Ala Gly Ala Leu Lys Asp 50 55 60Arg Ala Pro Leu Leu Ala Arg Val Glu Glu Val Arg Val Glu Ser Arg 65 70 7580 Thr Pro Arg Ala Gly Gly Asp Leu Pro Thr Gly Phe Glu Ile Arg Glu 85 9095 Ser Arg Arg Thr Gly Arg Ala Asn Thr Ile Val Ala Pro Asp Ser His 100105 110 Val Cys Asp Asp Cys Leu Arg Glu Val Leu Asp Pro Thr Asp Arg Arg115 120 125 Phe Arg Tyr Pro Phe Thr Asn Cys Thr His Cys Gly Pro Arg TyrSer 130 135 140 Leu Ile Lys Gly Leu Pro Tyr Asp Arg Glu Lys Thr Thr MetAla Val 145 150 155 160 Phe Ala Met Cys Glu Asp Cys Arg Ala Glu Tyr ValAsn Pro Ala Asp 165 170 175 Arg Arg Tyr His Ala Gln Pro Thr Ala Cys ProArg Cys Gly Pro Lys 180 185 190 Val Ala Leu Ser Ala Pro Asp Gly Val ValAla Glu Ala Asp Glu Ala 195 200 205 Leu Thr Arg Thr Ile Arg Ala Leu GlyAsp Gly Gln Ile Val Ala Val 210 215 220 Lys Ser Val Gly Gly Phe His LeuAla Val Asn Ala Arg Asp Ala Ala 225 230 235 240 Ala Val Ala Arg Leu ArgArg Arg Lys Lys Arg Asp Ser Lys Pro Phe 245 250 255 Ala Val Met Ala AlaGly Leu Asp Glu Thr Ala Ala Ile Ala His Ile 260 265 270 Thr Asp Ala GluArg Glu Leu Leu Arg Ser Pro Ala Arg Pro Ile Val 275 280 285 Leu Leu ArgLys Leu Pro Gly Ala Leu Pro Glu Glu Val Ala Pro Arg 290 295 300 Asn ProAsn Leu Gly Val Met Leu Pro Ser Ala Pro Leu His Gln Leu 305 310 315 320Leu Leu Asp Arg Pro Glu Ser Asp Val Leu Val Met Thr Ser Gly Asn 325 330335 Ile Ser Gly Gln Pro Ile Ala Tyr Arg Asn Glu Asp Ala Leu Ala Glu 340345 350 Leu Phe Glu Val Ala Asp Leu Ile Leu His His Asp Arg Asp Ile His355 360 365 Val Arg Val Asp Asp Ser Val Val Arg Ser Ser Val His Pro AlaLeu 370 375 380 Asp Glu Pro Ile Ile Thr Phe Leu Arg Arg Ser Arg Gly TyrAla Pro 385 390 395 400 Tyr Pro Val Lys Val Asp Asp Ile Ala Gly Pro ValIle Ala Tyr Gly 405 410 415 Ala Glu Leu Lys Thr Thr Val Ala Leu Gly AspGly Asn Glu Val Tyr 420 425 430 Val Ser Gln His Ile Gly Asp Leu Lys AsnAsp Glu Thr Tyr Ala Ser 435 440 445 His Arg Glu Ala Ala Ala His Leu AlaGlu Leu Tyr Glu Leu Lys Pro 450 455 460 Arg His Ala Ala Cys Asp Leu HisPro Ala Phe Lys Ala Arg Val Leu 465 470 475 480 Ala Gly Thr Asp Arg ProAsp Asp Val Val Glu Val Gln His His His 485 490 495 Ala His Met Ala SerCys Met Ala Glu Asn Arg Leu Ser Gly Pro Thr 500 505 510 Ile Gly Val ValPhe Asp Gly Ala Gly Tyr Gly Glu Asp Gly Thr Ile 515 520 525 Trp Gly GlyGlu Phe Leu Val Gly Asp Phe Ala Ser Val Arg Arg Ala 530 535 540 Ala HisLeu Arg Pro Val Pro Leu Leu Gly Gly Asp Lys Ala Val Ala 545 550 555 560Glu Pro Val Arg Thr Gly Tyr Ala Leu Ala Leu Gln Ala Leu Gly Ser 565 570575 Pro Glu Arg Ala Val Ala Ala Phe Pro Ala Leu Glu Ile Leu Asp Asp 580585 590 Arg Lys Arg Thr Val Phe Ala Thr Met Ala Glu Arg Gly Ile Gln Ala595 600 605 Pro Pro Val Ser Ser Met Gly Arg Leu Phe Asp Ala Val Ala AlaLeu 610 615 620 Leu Gly Val Cys Ala Tyr Ala Glu Tyr Glu Ala His Gly ProIle Glu 625 630 635 640 Leu Glu Gly Leu Leu Gly Arg Asp Leu Ala Leu ThrAla Pro Tyr Ala 645 650 655 Phe Gly Leu Gln Gln Glu Gly Asp Thr Glu LeuVal Asp Pro Arg Pro 660 665 670 Leu Val Arg Ala Ile Ala Ala Asp Leu AlaArg Gly Thr Gly Val Thr 675 680 685 Asp Ile Ser Arg Arg Phe His Ser ThrVal Val Asp Met Val Val Lys 690 695 700 Arg Cys Ser Ala Leu Arg Glu ArgThr Gly Ile Arg Gln Val Val Leu 705 710 715 720 Ser Gly Gly Val Phe LeuAsn Glu Phe Leu Gln Val Asn Cys Leu Val 725 730 735 Glu Leu Arg Arg AlaGly Phe Asp Ala His Ala His Arg Leu Val Pro 740 745 750 Thr Asn Asp GlyGly Ile Ala Leu Gly Gln Val Met Val Ala Asn Ala 755 760 765 Arg Leu ArgAla Ser Pro Asp Ser Thr Asp Ser Pro Ala Thr Pro Asn 770 775 780 Pro GlyThr Asn Glu Cys Arg 785 790 33 447 PRT Streptomyces atroolivaceus 33 MetArg Thr Ala Asp Leu Thr Leu Asp Thr Val Val Arg Leu Leu Ser 1 5 10 15Glu Asn Glu Ala Ser Ala Arg Gln Thr Leu Ser Met Val Pro Ser Glu 20 25 30Thr Ser Met Ser Gly Leu Ala Lys Leu Pro Met Leu Leu Asp Pro Tyr 35 40 45His Arg Tyr Phe Phe Asn Glu Gly Asp Asp Pro Asp Arg Trp His Phe 50 55 60Arg Gly Ala Gln Arg Leu Arg Asp Leu Glu Met Glu Leu Thr Ile Pro 65 70 7580 Leu Leu Gln Glu Leu Gly Arg Ala Ser Tyr Ala Ser Val Arg Pro Leu 85 9095 Ser Gly Leu Asn Gly Met Thr Leu Val Leu Gly Ala Leu Gly Gly Glu 100105 110 Pro Gly Ser Thr Val Val Thr Val Ser Pro Glu Gln Gly Gly His Tyr115 120 125 Ala Thr Pro Gln Val Ala Gly Arg Leu Gly Leu His Ala Glu PheLeu 130 135 140 Arg Gly Pro Asp Pro His Ser Leu Asp Leu Glu His Ala AlaGlu Leu 145 150 155 160 Leu Ala Arg Val Arg Pro Ser Leu Val Tyr Val AspGln Ser His Cys 165 170 175 Leu Phe Pro Val Asp Val Lys Ser Leu Val GluThr Val Arg Glu Ala 180 185 190 Ser Pro Gly Thr Leu Val His Val Asp AlaSer His Trp Leu Gly Leu 195 200 205 Val Leu Gly Asp Ala Phe Pro Asn ProLeu Asp Gln Gly Ala Asp Ser 210 215 220 Trp Gly Gly Ser Thr His Lys ThrPhe Pro Gly Pro Gln Lys Ala Val 225 230 235 240 Val Leu Thr Arg Asp ProGln Val Glu Gln Leu Ile Arg Asp Ala Gln 245 250 255 Asp Phe Leu Ile SerAsn His His Phe Ala Ala Thr Ile Ala Leu Gly 260 265 270 Ile Ser Leu LeuGlu Phe Arg Glu Phe Gly Pro Ala Tyr Thr Arg Ala 275 280 285 Val Leu GluHis Thr Gly Arg Phe Gly Arg Leu Leu Thr Glu Arg Gly 290 295 300 Leu ThrVal Val Ala Ala Asp Arg Gly Tyr Ser Ala Gly His Gln Leu 305 310 315 320Trp Leu Asp Thr Glu Ala Asp Gly Ile Ala Pro Lys Asp Ala Ala Ala 325 330335 Arg Leu Ser Ala Ala Ser Leu Lys Val Asn Phe Met Ala Gly Leu Pro 340345 350 Gly Phe Thr Gly Gln Gly Val Arg Ile Gly Leu Asn Glu Ala Thr Tyr355 360 365 Gln Gly Leu Ser Gly Asp Asp Ile Asp Glu Leu Ala Asp Ile PheVal 370 375 380 Ala Ala Val Arg Asp Thr Glu Pro Ala Ala Leu Leu Ala GluArg Thr 385 390 395 400 Ala Ala Leu Arg Thr Arg Thr Pro Phe Gly Ala CysAla Ala Asp Gly 405 410 415 Ser Pro Leu Leu Ala Thr Ala Leu Asp Leu CysGlu Gly Ala Leu Arg 420 425 430 Arg Thr Gly Ala Ala Ala Gly Arg Pro LeuThr Glu Ala Ser Arg 435 440 445 34 238 PRT Streptomyces atroolivaceus 34Met Ser Arg Pro Val Ile Gly Ile Ala Ser Tyr Arg Asp Arg Ala Arg 1 5 1015 Trp Asn Ile Trp Asp Thr Asp Ala Thr Val Leu Gln Gln Gly Tyr Val 20 2530 Asp Gly Val Arg Arg Ala Gly Gly Arg Ala Val Val Leu Pro Pro Asp 35 4045 Asp Thr Asp Ala Asp Val Val Ala Arg Leu Asp Gly Leu Leu Leu Pro 50 5560 Gly Gly Ala Asp Val Asp Pro Ala Arg Tyr Gly Ala Ala Arg His Pro 65 7075 80 Arg Thr Asp Thr Pro Ser Ala Asp Arg Asp Ala Gly Glu Leu Leu Leu 8590 95 Leu Glu Ala Ala Leu Ala Ala Gly Leu Pro Val Leu Gly Val Cys Arg100 105 110 Gly Leu Gln Leu Leu Ala Leu Ala Tyr Gly Gly Thr Leu His GlnHis 115 120 125 Leu Pro Asp Leu Val Gly His Ser Gly His Cys Pro Ala GluGly Val 130 135 140 Phe Gly Arg His Pro Val His Leu Val Pro Gly Ser ArgAla Ala Ala 145 150 155 160 Leu Tyr Gly Glu Leu Ala Glu Val Asn Ser HisHis His Gln Ala Val 165 170 175 Leu Asp Pro Gly Gly Leu His Val Thr GlyArg Gly Glu Asp Gly Val 180 185 190 Val Glu Ala Ala Glu Asp Pro Ser ArgPro Phe Val Leu Gly Val Gln 195 200 205 Trp His Pro Glu Val Ser Asp AspArg Lys Leu Phe Ala Ala Phe Val 210 215 220 Ala Ala Cys Thr Thr Arg SerLeu His Thr Ala Ala Ser Val 225 230 235 35 1745 PRT Streptomycesatroolivaceus 35 Met His Thr His Val Arg Pro Val Gly Gly Thr Leu Val AspIle Leu 1 5 10 15 Arg Asp Arg Ala Asp Ala Thr Pro His Arg Thr Ala LeuGlu Phe Ile 20 25 30 Arg Asp Asp Gly Ser Glu His Ala Val Gly Tyr Ala AlaLeu Asp Arg 35 40 45 Arg Val Arg Ala Val Ala Ala Ala Leu Thr Glu Arg GlyLeu Arg Gly 50 55 60 Glu Arg Val Leu Leu Leu Phe Pro Pro Gly Glu Asn TyrVal Thr Ala 65 70 75 80 Phe Leu Gly Cys Leu Tyr Ala Gly Ala Val Ala ValPro Val Tyr Val 85 90 95 Pro Thr Gly Lys Arg Gly Leu Ser Ala Val Leu AlaThr Gly Ala Asp 100 105 110 Ala Gly Ala Val Leu Ala Leu Ser Ser Arg GluVal Thr Asp Ala Ile 115 120 125 Thr Ala Ser Tyr Pro Glu Leu Thr Thr SerGly Leu Ile Gln Trp Leu 130 135 140 Val Thr Glu Asp Ile Ala Asp Ala AlaAla Asp Asp Trp Pro Gly Thr 145 150 155 160 Gly Pro Ala Pro Ala Asp LeuAla Phe Leu Gln Tyr Thr Ser Gly Ser 165 170 175 Thr Gly Thr Pro Lys GlyVal Met Val Arg His Asp Asn Leu Val His 180 185 190 Asn Ser Ala Ser IleSer Thr Ala Leu Gly Val Gly Pro Asp Ser Arg 195 200 205 Gly Val Ser TrpLeu Pro Pro Tyr His Asp Met Gly Leu Ile Gly Gly 210 215 220 Ile Leu GlnPro Leu Tyr Ala Gly Phe Pro Cys Thr Leu Ile Ser Pro 225 230 235 240 MetAla Phe Val Arg Ser Pro Tyr Arg Trp Leu Asp Ala Ile Ser Arg 245 250 255His Arg Ala Thr Val Ser Ala Ala Pro Asp Phe Ala Tyr Gly Glu Cys 260 265270 Val Arg Arg Ile Pro Glu Asp Lys Arg Ala Glu Leu Asp Leu Ser Ser 275280 285 Trp Gln His Ala Met Val Gly Ala Glu Pro Val Arg Pro Ala Thr Leu290 295 300 Asp Ala Phe Ala Arg Ala Phe Ala Asp Ser Gly Phe Asp Arg SerAla 305 310 315 320 Phe His Pro Cys Tyr Gly Leu Ala Glu Ala Thr Leu PheVal Thr Gly 325 330 335 Gly Ala Pro Glu Arg Gly Glu Pro Arg Val Leu ArgAla Asp Arg Arg 340 345 350 Glu Leu Glu Leu Gly Gln Ala Val Ser Ala IleAla Gly Lys Glu Asn 355 360 365 Lys Glu Asn Ala Ala Val Leu Thr Gly CysGly Arg Ser Arg Ser Glu 370 375 380 Asp Val Val Val Val Val Asp Pro AlaGlu Gly Arg Ala Cys Pro Pro 385 390 395 400 Gly Gly Thr Gly Glu Val TrpVal Ser Gly Pro Thr Val Thr Ala Gly 405 410 415 Tyr Trp Gly Arg Pro GluLeu Thr Asp Glu Val Phe Arg Ala Thr Leu 420 425 430 Glu Ser His Pro GlyArg His Phe Leu Arg Thr Gly Asp Leu Gly Phe 435 440 445 Ala Leu Gly AspGlu Leu Phe Val Thr Gly Arg Ala Lys Asp Leu Leu 450 455 460 Val Val ArgGly Arg Asn His Tyr Pro Gln Asp Ile Glu Gln Ser Ala 465 470 475 480 GluHis Ala His Pro Leu Leu Gln Pro Thr Arg Ala Ala Val Phe Ser 485 490 495His Asp Asp Gly Thr Glu Glu Arg Ala Val Leu Val His Glu Val Val 500 505510 Arg Gly Phe Asp Pro Glu Arg Ala Pro Glu Val Ile Thr Ala Val Arg 515520 525 Lys Ala Val Thr Ala Glu His Gly Leu Ser Leu His Glu Val Val Leu530 535 540 Val Arg Pro Gly Ala Ile Pro Arg Thr Thr Ser Gly Lys Val ArgArg 545 550 555 560 Ser Thr Cys Arg Glu Arg Trp Leu Asp Gly Thr Leu GlnPro Ile Ala 565 570 575 Thr Gly Gly Ala Asp Thr Ala Pro Asp Ala Gly GlyGly Asp Met Pro 580 585 590 Pro Val Ser Arg Ala Val Ala Glu Val Val AlaLeu Glu Leu Asp Val 595 600 605 Pro Ala Glu Gln Leu Pro Pro Gly Val ProLeu Val Val Leu Asp Leu 610 615 620 Asp Ser Leu Arg Ala Val Arg Leu AlaGlu Ala Met Arg Glu Arg Phe 625 630 635 640 Gly Leu Arg Val Ser Pro AlaGlu Leu Leu Asp Gly Leu Thr Leu Ala 645 650 655 Gly Leu Ser Ala Leu ValAsp Ala Val Asp Asp Val Ala Glu Ala His 660 665 670 Val Gly Ala Asp AspThr Glu Ala Ala Asp Ala Pro Ala Ser Arg Ala 675 680 685 Gln Glu Trp MetTrp Leu Leu Asp Ala Met Gly Gly Gly Ala Ala Tyr 690 695 700 His Val AlaGly Gly Ile Asp Leu His Gly Pro Val Asp Pro Glu Leu 705 710 715 720 LeuArg Ala Ser Leu Glu Asp Leu Val Ala Arg His Pro Ala Leu His 725 730 735Ser Gly Phe Thr Ser Asp Pro Asp Gly Thr Leu Arg Arg Thr Pro Leu 740 745750 Ala Pro Arg Ser Phe Glu Leu Pro Val Val Glu Pro Ser Ala Asp Arg 755760 765 Asp Ala Arg Ala Val Ala Ala Glu Leu Ala Arg Glu Pro Phe Asp Leu770 775 780 Ala Ala Gly Pro Leu Leu Arg Ala Val Leu Val Arg Ala Gly GluHis 785 790 795 800 Glu Trp Thr Leu Ala Val Ala Ala His His Ile Ala ValAsp Gly Trp 805 810 815 Ser Leu Gly Leu Leu Leu Arg Glu Leu Gly Ile CysTyr Arg Asp Arg 820 825 830 Leu Ala Gly Arg Ser Pro Ser Arg Thr Val ValThr Val Pro Ala Pro 835 840 845 Lys Ser Ala Gly Asp Asp Ala Glu Ala AlaAla Phe Trp Gln Glu Thr 850 855 860 Leu Ala Gly Ala Gln Ala Val Ser LeuPro Leu Asp Pro Ala Pro His 865 870 875 880 Pro Ser Pro Phe Lys Gly SerAla Leu Pro Phe Thr Leu Pro Ala Pro 885 890 895 Val Val Ala Arg Leu LysArg Tyr Gly Ala Lys Arg Arg Ser Thr Leu 900 905 910 Phe Met Val Ile LeuThr Gly Leu Ser Ala Val Leu Ser Arg Trp Thr 915 920 925 Gly Gln Gln AspLeu Val Val Gly Ala Val Thr Ala Gly Arg His Arg 930 935 940 Pro Gly SerGly Asp Leu Val Gly Leu Phe Val Asn Thr Leu Pro Leu 945 950 955 960 ArgVal Glu Ala Ser Gly Asp Pro Ala Phe Gly Glu Leu Leu Ser Arg 965 970 975Val Arg Ser Ser Cys Leu Ala Ala Tyr Arg His Gln Asp Leu Pro Leu 980 985990 Glu Glu Ile Val Arg Leu Thr Gly Ala Asp Arg Ala Asp Gly Arg Ala 9951000 1005 Pro Leu Val Arg Val Ala Leu Ala Leu Gln Asn Leu Pro Val Ala1010 1015 1020 Pro Trp Gln Ser Gly Glu Val Arg Ala Glu Pro Phe Glu LeuPro 1025 1030 1035 Ser Pro Gly Ala Gln Phe Glu Leu Ser Leu His Leu ThrGlu Glu 1040 1045 1050 Ser Asp Gly Ser Leu Thr Gly His Ala Val His AlaSer Asp Leu 1055 1060 1065 Ile Ser Ala Pro Ser Ala Arg Arg Ile Leu GlnAla Leu Thr Leu 1070 1075 1080 Ala Leu Glu Ala Val Pro Gly Pro Pro GlyAla Ala Ala Thr Asp 1085 1090 1095 Leu Pro Met Leu Ala Pro His Glu LeuAsp Arg Val Val Ser Glu 1100 1105 1110 Phe Ser Gly Ser Asp Thr Pro LeuAsp Asp Gly Leu Val His Ala 1115 1120 1125 Gly Phe Glu Arg Gln Ala ArgLeu Thr Pro Asn Ala Pro Ala Val 1130 1135 1140 Val Trp Asp Gly Gly ThrLeu Ser Tyr Ala Gln Leu Asp Ala Arg 1145 1150 1155 Ala Asn Arg Leu AlaArg Arg Leu Arg Ser Leu Gly Ala Ala Pro 1160 1165 1170 Asp Gln Val ValAla Val His Leu Pro Arg Ser Pro Glu Leu Val 1175 1180 1185 Val Ala LeuLeu Ala Val Val Lys Ala Gly Gly Ala Tyr Leu Pro 1190 1195 1200 Leu AspPro Asp His Pro Arg Ala Arg Leu Ala Leu Gln Leu Ala 1205 1210 1215 AspVal Thr Pro Ser Leu Leu Val Thr Ser Arg Asp Ala Ala His 1220 1225 1230Gly Arg Ala Gly Glu Leu Ala Ala Gly Ser Pro Gly Leu Thr Leu 1235 12401245 Val Glu Ile Asp Asp Pro Ala Leu Ala Glu Leu Pro Asp Gly Pro 12501255 1260 Leu Asp Glu Val Ala Arg Pro Asp Asn Leu Leu Tyr Val Leu His1265 1270 1275 Thr Ser Gly Thr Thr Gly Thr Pro Lys Gly Val Met Asn ArgHis 1280 1285 1290 Ala Gly Val Ala Asn Arg Met Ala Trp Met Gln Glu LysTyr Pro 1295 1300 1305 Leu Val Ala Gly Glu Arg Val Leu His Lys Thr ProVal Gly Phe 1310 1315 1320 Asp Val Ser Gly Trp Glu Ile Trp Trp Pro LeuGly His Gly Ala 1325 1330 1335 Thr Ile Val Leu Ala Arg Pro Gly Gly HisGln Asp Pro Gly Tyr 1340 1345 1350 Leu Val Arg Ser Val Val Asp His AlaVal Thr Thr Cys His Phe 1355 1360 1365 Val Pro Ser Met Leu Arg Val PheLeu Asp Glu Thr Thr Ala Ala 1370 1375 1380 Thr Ala Gly Gly Thr Leu ArgArg Val Val Cys Ser Gly Glu Ala 1385 1390 1395 Leu Pro Pro Asp Val AlaAla Arg Phe His Arg Leu Leu Pro Gly 1400 1405 1410 Val Glu Leu His AsnLeu Tyr Gly Pro Thr Glu Ala Ala Ile Asp 1415 1420 1425 Val Thr Ala GluGln Val Glu Pro Gly Ala Thr Asp Arg Pro Arg 1430 1435 1440 Leu Pro IleGly Arg Pro Val Pro Gly Val Arg Leu Tyr Val Leu 1445 1450 1455 Asp GlyArg Gly Asn Pro Ala Pro Val Gly Val Pro Gly Glu Leu 1460 1465 1470 HisIle Gly Gly Pro Gln Val Ala Arg Gly Tyr Phe Gly Arg Pro 1475 1480 1485Ser Leu Thr Ala Ala Ala Phe Val Pro Asp Pro Phe Gly Ser Gly 1490 14951500 Gly Arg Leu Tyr Arg Thr Gly Asp Arg Ala Cys Trp Thr Glu Asp 15051510 1515 Gly Thr Leu Asp Tyr Leu Gly Arg Ile Asp His Gln Val Lys Ile1520 1525 1530 Arg Gly Gln Arg Val Glu Pro Gly Glu Ser Glu Ala Val LeuAla 1535 1540 1545 Arg His Pro Ala Val Gly Asp Ala Val Val Val Pro ArgArg Gln 1550 1555 1560 Gly Gly Glu Thr Tyr Leu Ala Ala Tyr Leu Val AlaVal Ala Gly 1565 1570 1575 Ala Glu Gln Pro Gly Pro Ala His Leu Arg GluHis Leu Ser Leu 1580 1585 1590 Ser Leu Pro Pro Ala Met Ile Pro Ser AlaTyr Thr Trp Leu Asp 1595 1600 1605 Ala Leu Pro Val Gly Ala Asn Gly LysLeu Asp Arg Asp Ala Leu 1610 1615 1620 Pro Glu Pro Asp Gln Asn Leu AlaGly Gly Glu His Val Ala Pro 1625 1630 1635 Arg Asp Ala Ala Glu Gln ArgVal Ala Glu Val Trp Cys Glu Val 1640 1645 1650 Leu Gly Leu Glu Glu ValSer Val Thr Ala Asp Phe Phe Ala Leu 1655 1660 1665 Gly Gly His Ser LeuGln Ala Thr Arg Ile Ala Leu Arg Leu Arg 1670 1675 1680 Glu Ala Phe GlyAla Glu Phe Ser Val Ala Asp Leu Leu Thr Gly 1685 1690 1695 Val Pro ThrVal Glu Gln Thr Ala Arg Leu Leu Thr Gln Arg Gln 1700 1705 1710 Val AlaGlu Ser Asp Pro Glu Glu Leu Ala Arg Leu Leu Asp Gln 1715 1720 1725 LeuSer Gly Leu Ser Asp Asp Glu Val Ala Asp Leu Leu Gln Gln 1730 1735 1740Gln Gly 1745 36 462 PRT Streptomyces atroolivaceus 36 Met Ala Val ThrLeu Lys Asp Asp Phe Ala Glu Arg Ala Arg Leu Glu 1 5 10 15 Ala Arg LeuLeu Ala Arg Arg Arg Gly Gly Ala Ala Leu Pro Leu Leu 20 25 30 Asp Arg ProAla Gly Ala Glu Trp His Phe Ala Ala Ser Leu Gly Gln 35 40 45 Glu Gly MetTrp Pro Ser Leu Ala Thr Thr Ala Arg Gln Pro Leu Ile 50 55 60 Val Gly GlyLeu Arg Val Ser Gly Pro Leu Asp Thr Gly Ala Leu Arg 65 70 75 80 Glu AlaTrp Gln Ala Leu Ala Glu Arg His Glu Thr Leu Arg Ser Ala 85 90 95 Tyr ArgArg Val Asp Gly Ala Leu Thr Gln Val Val Pro Val Arg Ala 100 105 110 ArgPro Val Met Asp Val Ala Leu Ala Asp Pro Ala Ala Ala Ala Ser 115 120 125Val Ala Asp Asp Glu Leu Ala Arg Pro Phe Asp Phe Gly Arg Gly Pro 130 135140 Leu Ala Arg Leu Arg Val Leu Arg Arg Ala Ser Gly Glu His Leu Val 145150 155 160 Val Val Cys Leu His His Leu Val Gly Asp Ala Arg Ala Leu GluVal 165 170 175 Val Thr Thr Glu Leu Thr Ala Leu Tyr Ala Gly Ala Val ThrGly Ser 180 185 190 Pro Val Gln Leu Pro Glu Leu Pro Ala Gln Tyr Ala AspPhe Ala Ala 195 200 205 Trp His Arg Ser Arg Val Glu Gly Glu His Gly GlyArg Thr Val Ala 210 215 220 Tyr Trp Leu Asp Arg Leu Ala Gly Ala Glu ProAla Ala Leu Pro Cys 225 230 235 240 Asp Arg Ser Gln Asp Pro Ser Asp IleAla Gly His Thr Arg Ser Phe 245 250 255 Pro Leu Pro Pro Glu Leu Phe AlaArg Leu Arg Glu Val Ala Ala Arg 260 265 270 His Arg Thr Thr Phe Tyr ValVal Gly Leu Ala Ala Phe Gln Ile Leu 275 280 285 Leu Ala Arg Trp Ser GlyGlu Arg Asp Ile Cys Val Arg Ala Pro Val 290 295 300 Ser Tyr Arg Asp ArgSer Glu Val His Gly Leu Val Gly Asp Phe Ser 305 310 315 320 Asn Asp ValVal Val Arg Ala Asp Leu Ser Gly His Val Thr Leu Ala 325 330 335 Glu AlaVal Glu Gln Val Arg Ala Ser Thr Ala Asp Asp Phe Ala His 340 345 350 HisAsp Val Pro Pro His Val Val Ala Ser Arg Met Ala Asp Pro Thr 355 360 365Leu Leu Glu Arg Leu Phe His Val Gln Phe Thr Ala Glu Arg Glu His 370 375380 Asp Leu Gly Ala Ala Arg Gln Leu Gly Pro Ile Arg Ala Glu Pro Phe 385390 395 400 Ser Pro Thr Ala Glu Ser Val Ala Arg Pro Leu Ser Val Arg LeuArg 405 410 415 His Asp Asp His Gly Gly Arg Val Ile Val Ala Tyr Arg SerGly Leu 420 425 430 Phe Ser Asp Glu Arg Val Arg Ala Leu Gln Arg Asp TyrPhe Ala Leu 435 440 445 Phe Glu Glu Ile Ala Ala His Pro Gly Gln Pro AlaVal Ser 450 455 460 37 399 PRT Streptomyces atroolivaceus 37 Met Ser AlaThr Arg Arg Val His Ile Tyr Pro Phe Glu Gly Glu Val 1 5 10 15 Asp GlyLeu Glu Ile His Pro Lys Phe Ala Glu Leu Arg Glu Thr Asp 20 25 30 Pro LeuAla Arg Val Arg Leu Pro Tyr Gly Gly Glu Gly Trp Met Val 35 40 45 Thr ArgTyr Asp Asp Val Arg Ala Ala Asn Ser Asp Pro Arg Phe Ser 50 55 60 Arg AlaGln Ile Gly Glu Asp Thr Pro Arg Thr Thr Pro Leu Ala Arg 65 70 75 80 ArgSer Asp Thr Ile Leu Ser Leu Asp Pro Pro Glu His Thr Arg Leu 85 90 95 ArgArg Leu Leu Ser Lys Ala Phe Thr Ala Arg Arg Met Gly Ala Met 100 105 110Gln Ser Trp Leu Glu Glu Leu Phe Ala Gly Leu Leu Asp Gly Val Glu 115 120125 Arg Thr Gly His Pro Ala Asp Ile Val Arg Asp Leu Ala Gln Pro Phe 130135 140 Thr Ile Ala Val Ile Cys Arg Leu Leu Gly Val Pro Tyr Glu Asp Arg145 150 155 160 Gly Arg Phe Gln His Trp Ser Glu Val Ile Met Ser Thr ThrAla Tyr 165 170 175 Ser Lys Glu Glu Ala Val Ser Ala Asp Ala Ser Ile ArgAla Tyr Leu 180 185 190 Ala Asp Leu Val Ser Ala Arg Arg Ala Ala Pro HisAsp Asp Leu Leu 195 200 205 Gly Val Leu Val Ser Ala Arg Asp Asp Asp AspArg Leu Thr Glu Asp 210 215 220 Glu Leu Ile Thr Phe Gly Val Thr Leu LeuVal Ala Gly His Glu Thr 225 230 235 240 Ser Ala His Gln Leu Gly Asn MetVal Tyr Ala Leu Leu Thr His Glu 245 250 255 Asp Gln Leu Ser Leu Leu ArgGlu Gln Pro Glu Leu Leu Pro Arg Ala 260 265 270 Val Glu Glu Leu Leu ArgPhe Val Pro Leu Gly Asn Gly Val Gly Asn 275 280 285 Ala Arg Ile Ala LeuGlu Asp Val Glu Leu Ser Gly Gly Thr Val Arg 290 295 300 Ala Gly Glu GlyVal Val Ala Ala Ala Val Asn Ala Asn Arg Asp Pro 305 310 315 320 Arg AlaPhe Asp Asp Pro Asp Arg Leu Asp Ile Thr Arg Glu Lys Asn 325 330 335 ProHis Leu Ala Phe Gly His Gly Ala His Tyr Cys Leu Gly Ala Gln 340 345 350Leu Ala Arg Met Glu Leu Arg Val Ala Ile Gly Gly Leu Leu Glu Arg 355 360365 Phe Pro Gly Leu Arg Leu Ala Val Pro Ala Asp Gln Val Glu Trp Lys 370375 380 Thr Gly Gly Leu Phe Arg Gly Pro Gln Arg Leu Pro Ile Ala Trp 385390 395 38 78 PRT Streptomyces atroolivaceus 38 Met Ala Arg Glu Gln AsnThr Arg Trp Arg Ile Glu Ala Asp His Gly 1 5 10 15 Ala Cys Met Gly ThrGly Ile Cys Ala Gly Val Ala Pro Asp Val Phe 20 25 30 Gln Val Val Asp GlyArg Ser Val Pro Pro Ala Gly Glu Val Pro Ala 35 40 45 Asp Glu Asn Val LeuAsp Ala Val Glu Ser Cys Pro Met Glu Ala Leu 50 55 60 Arg Val Arg Asp AlaVal Ser Gly Glu Val Leu Asp Leu Ser 65 70 75 39 115 PRT Streptomycesatroolivaceus 39 Met Lys Phe Ala Ile Val Ile Ile Glu Asn Ala Glu Ser ArgArg Gln 1 5 10 15 Ile Gln Glu Asp Arg Ala Thr Tyr Arg Lys Ala Ile GluGly Trp Met 20 25 30 Gly Gln Gln Ala Gln Ala Gly Ala Leu Val Gly Gly GluAla Phe Glu 35 40 45 Thr Glu Ser Leu Ala Pro Val Thr Val Arg Arg Glu AlaAsp Gly Thr 50 55 60 Arg Lys Ala Val Glu Gly Pro Phe Ala Gly Ala Asp GluThr Leu Gly 65 70 75 80 Gly Tyr Ile Leu Val Glu Ala Ala Asp Arg Asp AlaAla Val Glu Ile 85 90 95 Ala Lys Ser Trp Pro Asn Pro Glu Thr Leu Glu ValArg Pro Leu Trp 100 105 110 Val Ala Glu 115 40 438 PRT Streptomycesatroolivaceus 40 Met Thr Asp Gly Ala Ile His Thr His Ala Ala Gly Asp GlyAla Gly 1 5 10 15 Arg Leu Arg Pro Leu Leu Asp Ala Val His Asp Thr AlaCys Glu Val 20 25 30 Arg Ala Arg Arg Pro Asp Tyr Phe Glu His Arg Pro LeuArg Leu Pro 35 40 45 Ala Glu Leu Leu Ala His His Gly Thr Thr Pro Ala AlaLeu Ala Gln 50 55 60 Leu Ile Gly Asp Ser Tyr Glu Leu Thr Val Glu Pro ValThr Asp Pro 65 70 75 80 Ala Asp Cys Thr Ser Val Asp Arg Leu Ala Ala AlaArg Ala Gln His 85 90 95 Ala Gly Thr Thr Pro Gly Asp Ser Gly Ala Gln ArgAla Pro Gln Pro 100 105 110 Gly Glu Leu Ala Ala Val Ala Arg Glu Gln SerGlu Leu Arg Arg Ala 115 120 125 Ala Glu Arg Asp Ala Ala Arg Ile Gly AlaAla Ala Pro Leu Arg Val 130 135 140 Gly Val Leu Gly Gly Val Leu Pro TyrThr Ala Ala Gly Val Arg Glu 145 150 155 160 Glu Gly Glu Pro Pro Val ValLeu Val Asn Ala Leu Gly Met Gly Thr 165 170 175 Ala Leu Trp His Pro LeuMet Ala Ala Leu Ala Pro Thr Arg Arg Val 180 185 190 Leu Thr Trp Ala ProArg Gly Thr Ala Ala Gly Thr Arg Pro Met Arg 195 200 205 Leu Arg Asp GlnAla Asp Asp Leu Ala Ala Val Leu Ala Thr Glu Gly 210 215 220 Ala Arg AspCys His Leu Val Cys Trp Cys Ser Gly Pro Lys Thr Ala 225 230 235 240 MetGlu Phe Arg Arg Arg Arg Pro Glu Ser Val Arg Ser Met Val Phe 245 250 255Leu His Gly Ser Phe Arg His Thr Gly Asp Pro Ala Gly Leu Glu Pro 260 265270 Asp Thr Pro Tyr Glu Arg Asn Leu Glu Glu Leu Cys Arg Ala Val Val 275280 285 Ala Arg Pro Ala Leu Ala Ala Arg Leu Arg Thr Met Phe Thr Gly Gln290 295 300 Pro Ala Glu Met Ser Gly Asp Leu Thr Ala His Arg Phe Ala AspGlu 305 310 315 320 Val Leu Ala Leu Pro Ala Pro Ala Leu Gly Gly Glu LeuLeu Arg Pro 325 330 335 Phe Ser Asp Asp Ala Val Leu Val Ala Tyr Ala ArgGln Leu Leu Asp 340 345 350 Tyr Trp Ala Tyr Asp Gly Leu Ala His Ala AlaAsp Val Gln Val Pro 355 360 365 Val Leu Cys Val Ser Gly Glu Phe Asp ArgIle Ala Ser Pro Glu Arg 370 375 380 Leu Ala Leu Ala Ala Asp Arg Phe ProGly Ala Arg His Glu Arg Leu 385 390 395 400 Ala Gly Ala Thr His His SerMet Tyr Asp Arg Pro Arg Glu Val Ala 405 410 415 Arg Leu Leu Thr Asp PhePhe Thr Glu Ala Val Arg Pro Arg Ser Ile 420 425 430 Val Ser Gly Ala AlaArg 435 41 307 PRT Streptomyces atroolivaceus 41 Met Thr Asp Ala Ala SerGln Ala Gln Gly Ala Phe Leu Pro Ala His 1 5 10 15 Ser Gly Pro Leu GlySer Gly Ser Asp Leu Leu His Ser Gly Asn Ala 20 25 30 Gly Phe Ile Ile HisArg Ala Gly Gln Leu Asn His Ala Phe Arg Ala 35 40 45 Gln Gly Arg Gln PheAla Thr Asp Leu Val Gly His Leu Asn Lys Val 50 55 60 Val Glu Gly Val AlaThr Ile Val Val His Glu Glu Ile Leu Gly Thr 65 70 75 80 Ala Asp Arg LeuHis Trp Leu Ile His Met Lys Gln Pro Asn Asp Tyr 85 90 95 Ser Arg Phe LeuGlu Ile Ala Asp His Asp Arg Ser Phe Lys Glu Ile 100 105 110 Thr Glu AlaAsp Arg Ile Ala Ala Ala Glu Gly Ala Gly Asn Trp Glu 115 120 125 Arg MetPhe Val Glu Gly Ser Phe Gln Glu Arg Val Tyr Val Pro Gln 130 135 140 HisGly Leu Asp Glu His Asp Asp Asp His Asp His Asp His His Asp 145 150 155160 Glu Pro Ser Asp Thr Phe Val Pro Pro Ala Arg His Gln Thr Gly Leu 165170 175 Pro Asp Ser Arg Met Arg Ser Ser Val Asp Ser Gly Leu Thr Ile Ile180 185 190 Arg Thr Ala Gln Thr Ala Phe Arg Phe Arg Thr Glu Ala Arg GluPhe 195 200 205 Ala Phe Ala Trp Ala Ser Glu Val Asn Arg Ala Leu Gly GlyGlu Leu 210 215 220 Thr Val Tyr Leu Tyr Glu Glu Thr Phe Gly Gln Gln AspArg Ile His 225 230 235 240 Trp Met Ile His Leu Asp Ser Leu Asp Thr TyrArg Lys Leu Thr Glu 245 250 255 Leu Ser Arg His Asp Ala Asp Tyr Gln AlaLeu Phe Gly Arg Gln Phe 260 265 270 Val Pro Asp Phe Lys Gly Gly Gly GlyTrp Glu Gln Thr Phe Val Ser 275 280 285 Pro Thr Ile Gln Asp Thr Val LeuThr Pro Leu His Pro Gly Ala Pro 290 295 300 Ala Trp Phe 305 42 265 PRTStreptomyces atroolivaceus 42 Met Thr Ala Ile Gly Pro Thr His Arg GlyVal Arg Leu Thr Ala Glu 1 5 10 15 Pro His Val Leu Arg Ala Thr Leu ThrSer Pro Asp Gly Leu Asn Ser 20 25 30 Leu Ser Gly Ala Ala Leu Asp Ala LeuGly Ala Ala Leu Asp Arg Ala 35 40 45 Glu Ala Asp Pro Glu Cys Arg Val LeuLeu Leu Glu Gly Ser Gly Gly 50 55 60 Thr Phe Cys Thr Gly Leu Asp Phe GluGlu Ala Ala Gly Asp Pro Ala 65 70 75 80 Gly Gly Ala Ser Gln Ala Gly ArgGly Gly Ala Glu Phe Leu Ala Leu 85 90 95 Met Arg Arg Phe Gly Glu Thr ProLeu Ala Val Val Ala Cys Val Asp 100 105 110 Gly Arg Ala Ala Gly Gly GlyVal Gly Leu Ala Ala Ala Ala Asp Leu 115 120 125 Val Ile Ala Thr Glu ArgSer Glu Phe Ser Leu Pro Glu Ala Leu Trp 130 135 140 Gly Leu Val Pro CysCys Val Leu Pro Val Leu Val Arg Arg Thr Gly 145 150 155 160 Phe Gln ProAla Tyr Ala Met Ala Leu Ser Thr Gln Pro Val Ser Ala 165 170 175 Arg ArgAla Ala Asp Phe Arg Leu Val Asp Glu Val Val Pro Asp Pro 180 185 190 AspAla Ala Val Arg Arg Leu Leu Val Arg Leu Thr Arg Leu Asp Pro 195 200 205Ala Thr Ile Gly Glu Leu Lys Gln Tyr Phe Arg Ala Met Trp Phe Thr 210 215220 Thr Glu Asp Thr Asp Ala Phe Ala Leu Arg Glu Phe Thr Arg Leu Ile 225230 235 240 Asp Ser Pro Val Ala Arg Arg Arg Ile Thr Asp Tyr Thr Thr ThrArg 245 250 255 Arg Leu Pro Trp Glu Lys Pro Arg Pro 260 265 43 795 PRTStreptomyces atroolivaceus 43 Met Val Ala Leu Val Phe Pro Gly Gln GlySer Gln Arg Lys Gly Met 1 5 10 15 Gly Ala Asp Leu Phe Ala Arg Phe ProAsp Leu Thr Arg Gln Ala Asp 20 25 30 Thr Val Leu Gly His Ser Val Glu GluLeu Cys Arg Ser Ser Gly Asp 35 40 45 Gly Arg Leu Asp Arg Thr Glu Tyr AlaGln Pro Ala Leu Phe Val Val 50 55 60 Ser Ala Leu Ser Tyr Leu Ala Arg AspPro Gly Leu Pro Gln Pro Thr 65 70 75 80 Leu Leu Ala Gly His Ser Leu GlyGlu Tyr Gly Ala Leu Phe Ala Ala 85 90 95 Gly Cys Phe Asp Phe Ala Thr GlyVal Arg Leu Val Arg Glu Arg Gly 100 105 110 Ala Leu Met Gly Arg Ala GlnGly Gly Gly Met Leu Ala Val Leu Gly 115 120 125 Val Asp Gly Asp Glu ValGln Ala Leu Leu Ala Gly Thr Gly Ala Arg 130 135 140 Gln Val Asp Val AlaAsn Tyr Asn Thr Pro Thr Gln Thr Val Leu Ser 145 150 155 160 Gly Pro LeuAsp Glu Leu Arg Met Val Ser Ala Ala Leu Gly Gln Arg 165 170 175 Pro GlyVal Arg Cys Val Pro Val Arg Val Ser Ala Ala Phe His Ser 180 185 190 ArgHis Met Arg Pro Ala Ala Gln Glu Phe Ala Thr Phe Leu Thr Gly 195 200 205Phe Ser Phe Ala Asp Pro His Arg Thr Val Ile Ser Ser Val Thr Ala 210 215220 Arg Pro Tyr Gly Ala Gly Gln Val Ala Glu Leu Leu Ser Arg Gln Ile 225230 235 240 Glu Ser Pro Val Arg Trp Ser Glu Thr Met Ala Tyr Leu Arg GluArg 245 250 255 Gly Thr Thr Glu Leu Glu Glu Met Gly Pro Gly Lys Val LeuThr Gly 260 265 270 Leu Trp Lys Gln Gly Arg Ala Asp Gly Ala Lys Ala ArgAla Val Ala 275 280 285 Pro Ala Pro Val Ala Val Val Ala Gly Val Pro AlaAla Ala Ala Ala 290 295 300 Arg Ala Pro Ala Ser Pro Ala Pro Val Ala AlaArg Ala Ala Thr Ala 305 310 315 320 Ala Pro Ala Arg Pro Ser Asp Pro AlaPro Pro Ala Pro Arg Thr Ala 325 330 335 Pro Ser Pro Ala Ser Pro Val ProPro Ala Ser Val Ser Arg Gly Gln 340 345 350 Arg Ala Glu Glu Leu Gly SerAla Glu Phe Arg Gln Asp Tyr Gly Ile 355 360 365 Arg Tyr Ala Tyr Leu AlaGly Ala Met Phe Arg Gly Ile Ala Ser Ala 370 375 380 Glu Leu Val Ile ArgMet Gly Arg Ala Gly Leu Met Gly Phe Phe Gly 385 390 395 400 Ala Gly GlyLeu Gly Leu Asp Lys Val Glu Ser Ala Leu Val Arg Ile 405 410 415 Lys AspAla Leu Gly Pro Asp Gly Arg Tyr Gly Met Asn Leu Leu His 420 425 430 SerIle Asp Asp Pro Ala Tyr Glu His Ala Val Val Asp Leu Cys Leu 435 440 445Lys His Gly Val His Asp Val Glu Ala Ala Gly Phe Thr Gln Leu Thr 450 455460 Pro Ala Val Val Gln Phe Arg Phe Ser Gly Ala His Arg Asp Ala Ala 465470 475 480 Gly Arg Ala Val Ala Val Arg Arg Val Leu Ala Lys Val Ser ArgPro 485 490 495 Glu Val Ala Ala Ala Phe Met Ala Pro Ala Pro Ala Ala IleLeu Arg 500 505 510 Arg Leu Thr Ala Asp Gly Arg Leu Thr Pro Gln Glu AlaGlu Ile Ala 515 520 525 Ala Glu Leu Pro Val Gly Gln Asp Ile Cys Val GluAla Asp Ser Gly 530 535 540 Gly His Thr Asp Gly Gly Ala Ala Leu Thr LeuLeu Pro Ser Met Ile 545 550 555 560 Arg His Arg Asp Ala Ala Met Ala ArgHis Gly Tyr Gly Arg Arg Ile 565 570 575 Arg Ile Gly Ala Ala Gly Gly IleGly Ala Pro Glu Ala Val Ala Ala 580 585 590 Ala Phe Val Leu Gly Ala AspPhe Val Leu Thr Gly Ser Val Asn Gln 595 600 605 Cys Ser Pro Glu Ala GlyThr Ser Asp Ala Val Lys Asp Ile Leu Ala 610 615 620 Gly Leu Asp Val GlnAsp Thr Ala Tyr Ala Pro Ala Gly Asp Met Phe 625 630 635 640 Glu Ile GlyAla Arg Val Gln Val Val Arg Lys Gly Thr Leu Phe Ala 645 650 655 Ala ArgGly Asn Lys Leu Tyr Gln Leu Tyr Arg Ser His Asp Ser Trp 660 665 670 GluSer Ile Asp Ala Gly Thr Arg Arg Ser Val Glu Glu Thr Tyr Phe 675 680 685Lys Arg Pro Phe Ala Glu Val Trp Glu Glu Thr Arg Ala Tyr His Leu 690 695700 Gly Arg Gly Arg Asp Ala Glu Ile Glu Lys Ala Asp Arg Leu Pro Lys 705710 715 720 His Arg Met Ala Leu Ala Phe Arg Trp Tyr Phe Ala Arg Ser ValArg 725 730 735 Trp Gly Leu Glu Gly Glu Pro Thr Gln Lys Val Asn Tyr GlnIle Gln 740 745 750 Cys Gly Pro Ala Ile Gly Ala Phe Asn His Val Val ArgGly Thr Gly 755 760 765 Leu Glu Asp Trp Arg His Arg His Val Asp Leu IleAla Glu His Leu 770 775 780 Met Thr Gly Ala Ala Asp Val Leu Ala Arg Arg785 790 795 44 274 PRT Streptomyces atroolivaceus 44 Met Thr Thr Leu ThrPhe His Ser Thr Asn Ser Gly Val Ile Val Glu 1 5 10 15 Arg Thr Ala GlnLeu Lys Gln Ala His Arg Ala Glu Gly Arg Arg Ile 20 25 30 Ala Leu Glu GlnAla Ala Tyr Leu Asn Asp Lys Phe Ala Gly Gln Thr 35 40 45 Thr Val Thr ValHis Glu Glu Thr Phe Gly Val Arg Asp Arg Leu His 50 55 60 Trp Leu Val HisLeu Pro Lys Leu Ser Gly His Arg Glu Ile Ser Arg 65 70 75 80 Leu Leu AspGly Asp Glu Gly Pro Ala Ala Asp Ala Asp Trp His Gly 85 90 95 Met Phe ValAsp Gly Ser Phe Ser Glu Thr Ala Leu Ile Pro Gln His 100 105 110 Trp GlyMet Tyr Gly Thr Asp Glu Ala Leu Pro Glu Gly Thr Val Ile 115 120 125 AspAla Ala Ala Pro Asp Leu Arg Val Pro Pro Ala Gln Arg Gln Thr 130 135 140Ser Met Ser Pro Glu Arg Thr Leu Asn Ser Ser Gly Ala Gly Leu Met 145 150155 160 Ile His Arg Val Ala Gln Pro Lys Tyr Ala Phe Arg Ala Glu Ala Arg165 170 175 Leu Phe Ala Arg Arg Ile Thr Glu Ser Ile Asn Thr Arg Leu ProGly 180 185 190 Ile Ala Ser Ser Phe Leu Tyr Glu Glu Ala Phe Gly Pro AlaAsp Arg 195 200 205 Val His Trp Leu Ile His Met Lys Ser Glu Asp Thr TyrTyr Asp Leu 210 215 220 Ile Asp Met His Met Arg Met Asp Asp Ala Thr ArgAla Ile Tyr Leu 225 230 235 240 Asp Glu Ile Ile Ala Pro Glu Lys Gly GlyGly Thr Trp Asn Arg Leu 245 250 255 Phe Val Glu Glu Ser Met Gly Asp IleAla Phe Ser Pro Val Pro Asp 260 265 270 Ala Arg 45 4437 PRT Streptomycesatroolivaceus 45 Met Glu Pro Gly Thr Ser Pro Ser Gly Gln Gln Asp Ala SerGly Leu 1 5 10 15 Ala Ala Arg Val Thr Ala Val Met Val Arg Val Leu SerAla Ala Arg 20 25 30 Pro Asp Ala Gln Pro Val Thr Ala Ala Cys Glu Leu ArgGly Leu Gly 35 40 45 Leu Asp Ser Met Thr Ala Ala Arg Leu Trp Leu Ala ValGln Gly Glu 50 55 60 Cys Ala Ala Asp Val Pro Leu Gly Trp Leu Thr Glu AlaThr Thr Val 65 70 75 80 Gly Glu Tyr Ala Gln Arg Val Ala Asp His Ala SerGln Ala Val Pro 85 90 95 Val Gln Gly Ala Gly Ala Val Gly Ala Gln Val AlaAla Asp Pro Asp 100 105 110 Ala Leu His Glu Pro Phe Pro Leu Thr Pro LeuGln Glu Ala Tyr Leu 115 120 125 Ile Gly Lys Glu Pro Glu Leu Gln Ala AspAla Val Gly Cys His Leu 130 135 140 Tyr Arg Glu Phe Asp Val Pro Ala LeuAsp Thr Glu Arg Leu Arg Ala 145 150 155 160 Ala Trp Gln Arg Leu Val GluHis His Asp Ile Leu Arg Ala Thr Val 165 170 175 Thr Glu Asp Gly Arg GlnGln Ile Thr Ala Gln Ala Pro Arg Trp Asp 180 185 190 Leu Ala Val His GlySer Ala Thr Arg Ala Glu Phe Thr Glu Thr Ala 195 200 205 Thr Ala Val ArgAla Arg Met Ser His His Leu Phe Pro Ala Gly His 210 215 220 Cys Pro ProPhe Ala Ile Glu Val Thr Leu Gly Pro Asp Gly Thr Gly 225 230 235 240 ArgVal His Phe Gly Ile Asp Ala Ile Val Thr Asp Gly Gln Gly Leu 245 250 255Asp Leu Leu Thr Ala Gln Trp Glu Ala Cys Tyr Ala Asp Pro Ser His 260 265270 Leu Leu Pro Ala Pro Thr Ala Pro Leu Ser Val Arg Asp Cys Val Val 275280 285 Ala Leu Asp Ala Ala Arg Arg Thr Glu Ala His Arg Arg Asp Leu Asp290 295 300 His Trp Val Arg Arg Leu Arg Glu Leu Pro Gly Ala Pro Gly LeuPhe 305 310 315 320 Thr Ala Asp Ala Pro Glu Arg Gly Thr Gly Leu Ser CysVal Arg Arg 325 330 335 Ser Ser Arg Thr Ala Arg Leu Thr Ala Ala Glu TrpArg Ser Leu Arg 340 345 350 Ala Arg Ala Glu Glu Leu Ala Val Ser Pro ThrSer Leu Val Leu Thr 355 360 365 Val Phe Thr Glu Ala Leu Ala Arg His GlyAla His Glu Pro Phe Ser 370 375 380 Leu Val Val Thr Thr Ser Arg Arg ProGln Leu Pro Pro Glu Ala Asp 385 390 395 400 His Leu Val Gly Pro Phe ThrGly Thr Thr Phe Val Glu Ala Val Pro 405 410 415 Pro Gln Gln His Thr PheGlu Glu Ala Ala Arg Leu Thr His Glu Gly 420 425 430 Leu Trp Gln Ala LeuGlu His Ser Ala Val Cys Gly Val Ser Ala Gln 435 440 445 Arg Ala Leu ArgGly Gly Gly Pro Gly Pro Leu Pro Val Val Phe Thr 450 455 460 Ser Met LeuAsp Ala Ala Gly Arg Pro Arg Ala Arg Gly Phe Ala Ala 465 470 475 480 AlaPro Val Tyr Ala Val Ser Gln Thr Ser Gly Val Trp Leu Asp His 485 490 495Gln Met Trp Glu Gln Asp Gly Ala Leu His Leu Arg Trp Asp Thr Ala 500 505510 Asp Gly Cys Phe Ala Pro Gly Val Val Glu Ala Ala Phe Ala Ser Leu 515520 525 Cys Asn Gly Leu Arg Ala Leu Ala Val Ala Gly Pro Val Val Thr Arg530 535 540 Pro Leu Asn Asp Leu Gln Gln Ala Tyr Phe Val Ala Arg Ala AlaGly 545 550 555 560 Glu Pro Gly Pro Trp Arg Gly Cys Gln Val Val Val ProTyr Asp Thr 565 570 575 Asp Glu Arg Val Asp Pro Val Arg Leu Glu Ser AlaVal Val Arg Leu 580 585 590 Val Glu Ala Tyr Asp Val Leu Arg Ser Ala ValThr Gln Asp Gly Val 595 600 605 Val Glu Val Arg Ala Gly Ala Pro Arg ArgTrp Thr Val Pro Val Val 610 615 620 Ala Gly Gly Cys Pro Asp Glu Val ArgAsp Ala Met Ala Ala Ala Asn 625 630 635 640 Phe Pro Leu Gly Arg Tyr ProGln Phe Glu Val Arg Val Val Arg Gly 645 650 655 Asp Asp Gly Asp Thr ValLeu Met Ser Met Asp Leu Thr Leu Thr Asp 660 665 670 Ala Arg Gly Ile HisLeu Thr Gly Arg Glu Leu Met Arg Leu Tyr Ala 675 680 685 Asp Pro Ala AlaGlu Pro Arg Pro Ala Glu Ala Ala Arg Asp Ser Ala 690 695 700 Arg Asp AlaAsp Glu Gln Ala Arg Ser Arg Ala His Trp Gln Asp Arg 705 710 715 720 LeuArg Ala Leu Pro Pro Gly Val Pro Leu Pro Gly Pro Arg Asp Ala 725 730 735Asp Gly Pro Asp Arg Arg Val Arg Leu Ala Gly Ala Pro Leu Ala Leu 740 745750 Arg Pro Leu Thr Asp Arg Cys Ala Glu His Gly Leu Ser Leu Asp Ala 755760 765 Val Leu Leu Thr Ala Phe Thr Asp Val Leu Ala Arg Thr Tyr Gly Thr770 775 780 Asp Phe Ala Val Pro Val Val Arg Trp Asp His Gly Leu Asp ProGln 785 790 795 800 Arg Pro Gly Glu Phe Thr Ala Leu Ser Trp Leu Pro CysAla Pro Arg 805 810 815 Glu Leu Ser Phe Thr Ala Arg Ala Arg Thr Tyr GlnGlu Gly Leu Glu 820 825 830 Arg Asp Ala Asp Val Ser Gly Ser Gly Leu ProGlu Leu Arg Arg Ala 835 840 845 Val Ala Arg Ser Gly Gly Ala Gly Tyr ProVal Val Tyr Thr Cys Ala 850 855 860 Leu Asp Leu Thr Asp Arg Pro Leu ProGly Ser Val Arg Ala Gly Gln 865 870 875 880 Trp Leu Ser Cys Thr Pro AspVal Phe Leu Asp Cys Ile Thr Thr Val 885 890 895 Asp Ala Gly Gln Leu GlnLeu Ala Trp Asp Ala Val Asp Gly Arg Ala 900 905 910 Pro Gln Gly Gly TrpSer Glu Leu His Ala Glu Tyr Arg Arg Ser Val 915 920 925 Ala Arg Leu AlaAsp Asp Ala Ala Ala Trp Gln Glu Pro Ala Gly Gly 930 935 940 Asp Thr SerGly Ala Asp Asp Gly Glu Val Arg Gly Ala Glu Leu His 945 950 955 960 LysIle Leu His Glu Trp Asn Asp Thr Thr Arg Ala Phe Pro Asp Asp 965 970 975Arg Leu Met His Gln Leu Phe Glu Glu Gln Ala Ala Gln Gln Pro Arg 980 985990 Ala Glu Ala Leu Arg Trp Arg Gly Gly Gly Thr Met Thr Tyr Gln Glu 9951000 1005 Leu Asn Arg Arg Ala Asn Arg Ile Ala Ala Arg Leu Ala Ala Glu1010 1015 1020 Asp Val Gly Pro Glu Thr Val Val Ala Val Ser Val Pro ArgGly 1025 1030 1035 Pro Met Met Val Ala Val Val Leu Gly Ile Leu Lys AlaGly Gly 1040 1045 1050 Val Tyr Leu Pro Met Glu Pro His Leu Pro Ala GluArg Ala Ala 1055 1060 1065 Val Ile Leu Glu Glu Ala His Ala Glu Val ValVal Thr Thr Ala 1070 1075 1080 Asp Arg Glu Gly Trp Pro Val Pro Asp GlyTyr Ala Arg Val Cys 1085 1090 1095 Ala Asp Ala Ala Val Glu Gly Pro HisPro Ala Asp Ala Asp Asn 1100 1105 1110 Cys Pro Arg Pro Val Thr Gln ProHis Asn Thr Ala Tyr Ile Ile 1115 1120 1125 Phe Thr Ser Gly Ser Thr GlyArg Pro Lys Gly Val Ala Val Ala 1130 1135 1140 His Arg Pro Val Leu AsnLeu Ile Asn Trp Cys Arg Arg Thr Phe 1145 1150 1155 Gly Phe Gly Pro GlyAsp Met Gly Leu Cys Val Thr Ser Leu Gly 1160 1165 1170 Phe Asp Leu SerVal Phe Asp Val Phe Gly Leu Leu Gly Thr Gly 1175 1180 1185 Ala Ala LeuTyr Ile Ala Asp Ala Glu Gln Gln Arg Asp Pro Ala 1190 1195 1200 Leu LeuLeu Asp Val Leu Ile Glu Glu Pro Val Thr Phe Trp Asn 1205 1210 1215 SerAla Pro Thr Thr Leu Ala Gln Val Gly Pro Leu Leu Asp Thr 1220 1225 1230Val Gly Thr Ala Gly Thr Gly Asp Leu Arg Leu Val Phe Leu Ser 1235 12401245 Gly Asp Phe Thr Pro Leu Pro Leu Pro Asp Glu Val Arg Ala Val 12501255 1260 Phe Pro Arg Ala Asp Met Ile Ser Leu Gly Gly Ala Thr Glu Ala1265 1270 1275 Thr Val Trp Ser Asn Trp Phe Arg Ile Gly Ala Ile Asp ProAla 1280 1285 1290 Trp Arg Ser Ile Pro Tyr Gly Arg Pro Ile Asp Asn SerArg Tyr 1295 1300 1305 His Val Leu Asp Glu Ala Leu Arg Pro Cys Pro ValGly Val Glu 1310 1315 1320 Gly Asp Leu Tyr Ile Gly Gly Glu Cys Leu AlaLeu Gly Tyr Val 1325 1330 1335 Asn Gln Pro Glu Leu Thr Ala Asp Arg PheIle Pro Asp Pro Phe 1340 1345 1350 His Glu Asp Pro Gln Glu Arg Leu TyrLys Thr Gly Asp Arg Ala 1355 1360 1365 Leu Tyr Tyr Pro Asp Gly Asn LeuSer Phe Gln Gly Arg Ala Asp 1370 1375 1380 Gly Gln Val Lys Val Arg GlyPhe Arg Val Glu Leu Ala Glu Ile 1385 1390 1395 Glu His Arg Leu Arg AlaHis Asp Gly Val Lys Asp Ala Val Val 1400 1405 1410 Leu Ala Arg Glu AspGly Cys Gly Asp Arg Thr Leu Val Ala Tyr 1415 1420 1425 Leu Val Ala LeuPro Gly Ser Ala Pro Ser Gly Arg Glu Leu Arg 1430 1435 1440 Gly Phe AlaGly Gln Thr Leu Pro Glu Tyr Met Val Pro Asn Phe 1445 1450 1455 Ile GlyPhe Leu Ala Gly Phe Pro Ala Thr Ala Asn Gly Lys Leu 1460 1465 1470 AspArg Ala Ala Leu Pro Trp Pro Leu Ala Lys Ala His Leu Thr 1475 1480 1485Pro Pro Asp Arg Ser Ala Asp Ala Asp Pro Val Gly Ser Glu Ala 1490 14951500 Glu Ala Ala Val Pro Val Lys Glu Ser Ala Ala Glu Gln Pro Ala 15051510 1515 Val Ala Gly Gly Ala Gly Pro Ser Val Ser Val Pro Ser Arg Asp1520 1525 1530 Glu Leu Cys Ala Glu Ile Ala Asp Leu Phe Ala Gln Ala LeuGly 1535 1540 1545 Val Glu Ser Val Asp Ala Asp Thr Asp Leu Trp Asp GlnGly Ala 1550 1555 1560 Thr Ser Phe Thr Met Val Arg Val Ser Gly Ser LeuGln Arg Ser 1565 1570 1575 Tyr Lys Gln Arg Phe Pro Val Ser Ala Leu LeuAsp Asn Pro Ser 1580 1585 1590 Val Ser Ala Ile Ala Gly Trp Val His AlaGln Leu Gly Gly Gly 1595 1600 1605 Ala Asp Ala Glu Ser Thr Ala Ala AlaGlu Ala Glu Thr Ala Thr 1610 1615 1620 Ser Val Asp Ala Glu Thr Thr AlaThr Thr Val Thr Gln Thr Thr 1625 1630 1635 Ala Ala Ser Asp Glu Arg ProAsp Ser Gly Pro Gly Pro Val Asp 1640 1645 1650 Phe Phe Ala Thr Glu GluArg Glu Arg Phe Lys Arg Gln His Trp 1655 1660 1665 Asn Arg Arg Pro AspGlu Pro Gly Leu Pro Glu Val Pro Leu Gly 1670 1675 1680 Asp Ala Arg PheGlu Asp Glu Leu His Ala Trp Arg Ala Ser Arg 1685 1690 1695 Arg Asp PheLeu Asp Gln Pro Val Pro His Arg Ser Phe Ser Arg 1700 1705 1710 Leu LeuGly Leu Leu Arg Glu Thr Thr Gly Ala Asp Gly Thr Gly 1715 1720 1725 AlaLeu Tyr Pro Ser Ala Gly Asp Thr Tyr Ser Val Gln Val Tyr 1730 1735 1740Leu His Leu Thr Pro Asp Ala Val Glu Gly Leu Asp Ala Gly Leu 1745 17501755 Tyr Tyr Tyr Asp Pro Ser Arg His Ser Leu Arg Leu Leu Arg Ser 17601765 1770 Gly Val Leu Pro Asp Arg Gly Ala His Phe Tyr Tyr Asn Arg Pro1775 1780 1785 Val Phe Asp Arg Ser Arg Phe Gly Ile Tyr Leu Phe Gly GlnArg 1790 1795 1800 His Gly Ile Glu Pro Leu Tyr Ala Glu Glu Ser Leu ArgTyr Leu 1805 1810 1815 Thr Leu Glu Ser Gly Tyr Met Ser Gln Leu Leu MetLeu Gly Gln 1820 1825 1830 Ala Ala His Gly Val Gly Leu Cys Pro Ile GlyAla Leu Asn Thr 1835 1840 1845 Glu Gln Leu Ser Gln Trp Leu Gly Leu AspGlu Gly His Val Phe 1850 1855 1860 Leu Gln Ala Phe Leu Gly Gly Ala AlaGlu His Pro Gln Arg Thr 1865 1870 1875 Ala Gly Gly Thr Val Pro Phe PheThr Glu Pro Thr Asp Ser Asp 1880 1885 1890 Gly Asn Ser Gly Ser Gly AspSer Ser Thr Val Ile Thr Asp Ala 1895 1900 1905 Val Ala Pro Val Ser AlaAla Ala Glu Asp Ala Asp Ala Glu Pro 1910 1915 1920 Pro Ala His Thr AlaGlu Pro Ala Ala Val Ile Gly Met Ala Gly 1925 1930 1935 Arg Leu Pro GlyAla Gly Asp Leu Asp Ala Phe Trp Asp Asn Leu 1940 1945 1950 Val Ser GlyArg Thr Ala Ile Gly Pro Ala Pro Ala Ser Arg Pro 1955 1960 1965 Glu ThrAla Pro Ser Gly Ala Arg Ala Thr Gly Gly Phe Leu Pro 1970 1975 1980 HisIle Asp Arg Phe Asp Ser Leu Leu Phe His Val Ser Pro Gln 1985 1990 1995Glu Ala Pro Ala Leu Asp Pro Gln Ala Arg Leu Met Leu Glu Ser 2000 20052010 Val Trp Gln Cys Leu Asp Asp Ala Gly His Thr Ala Asp Ser Leu 20152020 2025 Arg Arg Ser Ala Gly Arg Val Gly Val Phe Ile Gly Ser Met Trp2030 2035 2040 His Asp Tyr Arg Gln Gln Gly Ala Asp Arg Trp Asn Gly GlyAsp 2045 2050 2055 Ser Ala Glu Val Ala Ala Thr Ala Ser Asp Ile Ala AsnArg Val 2060 2065 2070 Ser His Phe Phe Asp Phe Arg Gly Pro Ser Leu AlaVal Asp Thr 2075 2080 2085 Ser Cys Ser Ser Ser Phe Ala Ala Leu His LeuAla Val Glu Ser 2090 2095 2100 Leu Arg Arg Gly Glu Cys Gly Ala Ala ValVal Gly Ala Val Asn 2105 2110 2115 Leu Leu Ala His Pro Tyr His Trp GlyLeu Leu Asp Gly Leu Glu 2120 2125 2130 Leu Leu Ala Ala Asp Ala Pro ProAla Ala Tyr Ala Ala Glu Gly 2135 2140 2145 Ser Gly Trp His Pro Gly GluGly Val Gly Val Leu Leu Leu Arg 2150 2155 2160 Pro Ala Asp Ala Ala ArgArg Ala Lys Asp Thr Val His Gly Leu 2165 2170 2175 Ile Glu Gly Thr ArgIle Gly His Ala Gly Arg Ala Pro Arg Tyr 2180 2185 2190 Gly Ala Pro HisThr Ala Ala Leu Ala Asp Ser Leu Ala Arg Ala 2195 2200 2205 Leu Ala AspAla Ser Val Ile Pro Asp Glu Val Asp Tyr Val Glu 2210 2215 2220 Cys AlaAla Ala Gly Ala Gly Ile Ala Asp Ala Ala Glu Leu Glu 2225 2230 2235 AlaLeu Gly Ser Val Leu Ala Arg Cys Ala Gly Ala Ser Pro Val 2240 2245 2250Pro Val Gly Thr Leu Lys Pro Asn Ile Gly His Leu Glu Ala Ala 2255 22602265 Ser Gly Leu Ser Gln Leu Ile Lys Val Leu Leu Gln Ile Arg His 22702275 2280 Gly Arg Ile Ala Pro Thr Leu Val Ser Gly Glu Leu Ser Pro Leu2285 2290 2295 Val Asp Trp Asp Gly Leu Pro Val Glu Leu Val Asp Thr ProArg 2300 2305 2310 Ala Leu Thr Pro Arg Ala Ala Asp Gly Arg Ala Thr ValLeu Val 2315 2320 2325 Asn Ala Val Gly Ala Thr Gly Ser Tyr Gly His ValVal Val Arg 2330 2335 2340 Ala Pro His Ala His Gly Thr Gly Pro Ala AlaGln Asp Gly Leu 2345 2350 2355 Ala Gly Ala Gly Ala Ala Pro Ser Ala SerGly Pro Arg Thr Val 2360 2365 2370 Val Leu Ser Ala Ala Ser Pro Glu GlyLeu Thr Ala Ala Ala Gly 2375 2380 2385 Arg Leu Arg Asp His Leu Ala GlyAla Gly Arg Ala Leu Cys Leu 2390 2395 2400 Asp Asp Val Ala Trp Thr LeuGln Thr Gly Arg Ala Ser Leu Gly 2405 2410 2415 His Arg Leu Thr Leu SerAla Asp Gly Leu Asp Gly Val Arg Ala 2420 2425 2430 Gly Leu Thr Ala PheLeu Asp Gly Arg Ala Cys Pro Gly Leu Ala 2435 2440 2445 Thr Ala Ala AlaAsp Pro Ala Leu Ala Gly Val Pro Ala Gly Ala 2450 2455 2460 Gln Asp LeuAla Arg Ala Ala Gly Asp Trp Leu Arg Gly His Ala 2465 2470 2475 Val AspPhe Ala Arg Leu Trp Ser Ala Pro Ala Arg Arg Val Pro 2480 2485 2490 LeuPro Val Gln Asp Phe Thr Val Leu Ala Gln Glu Arg His Trp 2495 2500 2505Leu Ala Ala Pro Ala Ala Arg Arg Pro Asp Gly Ala Ala Gly Ser 2510 25152520 Ala Pro Ala Ala Pro Glu Ser Gly Gln Ser Ala Pro Pro Ala Ser 25252530 2535 Pro Gln Val Gln Asp Asp Arg Ala Asp Arg Ala Gln Glu His Val2540 2545 2550 Ala Ala Cys Phe Ala Glu Val Ser Gly Ile Pro Ala Glu GlnLeu 2555 2560 2565 His Pro Arg Val Pro Leu Glu His Tyr Gly Leu Ser SerArg Leu 2570 2575 2580 Val Ala Arg Phe Asn Glu Arg Leu Arg Gln Asp ValGln Gly Val 2585 2590 2595 Ser Ser Thr Val Leu Phe Glu Tyr Pro Asp LeuAla Gly Val Ala 2600 2605 2610 Ala His Leu Ala Ala His His Glu Gly ProTrp Ser Ala Ala Pro 2615 2620 2625 Asp Thr Gln Pro Ser Pro Pro Val ProSer Pro Asp Pro Leu Pro 2630 2635 2640 Val Pro Arg Thr Pro Ala Ala AlaLeu Gly Glu Ser Ala Ala Ala 2645 2650 2655 Asp Gly Pro Glu Pro Ile AlaVal Ile Gly Ile Ala Gly Arg Tyr 2660 2665 2670 Pro Gly Ala Gly Asp LeuGlu Thr Phe Trp Ser Asn Leu Ala Glu 2675 2680 2685 Gly Val Asp Ser ValGly Pro Leu Pro Ala Glu Arg Ala Arg Asp 2690 2695 2700 Gly Trp Pro ThrGlu Gln Met Trp Gly Gly Phe Leu Asp Gly Val 2705 2710 2715 Asp Arg PheAsp Ala Leu Phe Phe Gly Ile Ala Pro Arg Asp Ala 2720 2725 2730 Gln LeuMet Asp Pro Gln Glu Arg Gln Phe Leu Gln Val Val Trp 2735 2740 2745 GluThr Leu Glu Asp Ala Gly Cys Thr Arg Ala Arg Ile Arg Glu 2750 2755 2760Gln Leu Gly Ser Asp Val Gly Val Phe Val Gly Thr Met Tyr Asn 2765 27702775 Glu Tyr Pro Phe Phe Gly Val Glu Arg Ser Leu Ala Gly Glu Ser 27802785 2790 Ala Asp Thr Gly Ser Ala Val Ala Gly Ile Ala Asn Arg Val Ser2795 2800 2805 Tyr Phe Leu Asp Leu His Gly Pro Ser Leu Ala Val Asp ThrMet 2810 2815 2820 Cys Ser Ser Ser Leu Thr Ala Leu His Leu Ala Val GluSer Leu 2825 2830 2835 Arg Arg Gly Glu Cys Ala Ala Ala Val Ala Gly GlyVal Asn Leu 2840 2845 2850 Ser Leu His Pro His Lys Phe Arg Gln Gln ThrArg Leu Lys Met 2855 2860 2865 Ser Ser Ser Asp His Arg Cys Arg Ser PheGly Ala Gly Gly Asp 2870 2875 2880 Gly Phe Val Pro Ala Glu Gly Val GlyAla Val Leu Leu Lys Pro 2885 2890 2895 Leu Ser Ala Ala Glu Ala Asp GlyAsp Arg Ile His Ala Val Ile 2900 2905 2910 Arg Gly Thr Ala Val Asn HisGly Gly Lys Thr Asn Gly Tyr Met 2915 2920 2925 Val Pro Asn Pro Val AlaGln Gly Asp Leu Val Arg Ala Ala Leu 2930 2935 2940 Arg Arg Ala Gly AlaAsp Pro Ala Thr Ile Gly Tyr Val Glu Ala 2945 2950 2955 His Gly Thr GlyThr Gln Leu Gly Asp Pro Val Glu Ile Asn Gly 2960 2965 2970 Leu Asn ArgAla Phe Ala Gly Ala Ser Val Ala Pro Ala Ser Arg 2975 2980 2985 Ala IleGly Ser Val Lys Ala Asn Ile Gly His Ala Glu Ala Ala 2990 2995 3000 AlaGly Ile Ala Gly Leu Thr Lys Val Val Leu Gln Leu Arg His 3005 3010 3015Arg His Leu Val Pro Ser Leu His Thr Glu Glu Leu Asn Asp Ala 3020 30253030 Val Asp Trp Ala Ser Ser Pro Phe Glu Val Val Arg Glu Gly Arg 30353040 3045 Pro Trp Ala Pro Leu Thr Gly Ala Asp Gly Ala Pro Leu Pro Arg3050 3055 3060 Arg Ala Gly Leu Ser Ala Phe Gly Ala Gly Gly Ala Asn AlaHis 3065 3070 3075 Val Val Val Glu Glu Tyr Val Pro Gly Thr Ala Pro GluPro Thr 3080 3085 3090 Glu Pro Gly Val Pro Gly Val Leu Glu Pro Gln LeuIle Val Leu 3095 3100 3105 Ser Ala His Asp Leu Gly Arg Leu Arg Ala LeuAla Gly Arg Leu 3110 3115 3120 Arg Asp Arg Leu Gly Arg Asp Asp Arg ProAla Pro Ala Leu Ala 3125 3130 3135 Asp Val Ala His Thr Leu Gln Ser GlyArg Glu Pro Leu Arg Glu 3140 3145 3150 Arg Val Ala Leu Val Ala Tyr AspVal Ala Gly Leu Cys Arg Ala 3155 3160 3165 Leu Asp Leu Phe Ala Ser GlyAsp Thr Gly Ala Trp Val His Gly 3170 3175 3180 Arg Thr Pro Gly Gly AlaLeu Pro Asp Gly Pro Lys Ala Val Leu 3185 3190 3195 Asp Ala Ala Ala AspArg Asp Ala Glu Leu Leu Arg Leu Gly Arg 3200 3205 3210 His Trp Thr GlyGly Gly Thr Val Asp Trp Pro Gly Leu His Pro 3215 3220 3225 Val Arg ArgArg Leu Val Ser Leu Pro Ser Tyr Pro Phe Ala Glu 3230 3235 3240 Asp ArgHis Trp Leu Pro Glu Pro Arg Thr Ala Ala Pro Ala Ala 3245 3250 3255 AlaAla Ala Thr Leu Thr Glu Pro Ser Gly Thr Thr Leu Tyr Gly 3260 3265 3270Arg Thr Trp Arg Ala Leu Pro Pro Leu Ala Ala Ala Ala Ala Pro 3275 32803285 Ala Pro Ser Gly Arg Val Leu Cys Val Phe Ser Ala Pro Gly Glu 32903295 3300 Pro Val Ala Arg Ala Leu Ala Ala Leu Leu Gly Pro Asp Arg Val3305 3310 3315 Thr Leu Val Arg Ala Gly Ala Asp Ala Gly Asn Gly Val ProGly 3320 3325 3330 Ile Thr Gly Ile Gly Asp Glu Ala Glu Ala Ala Ala PheAla Gln 3335 3340 3345 Gly Leu Arg Ala Asp Gly Pro Asp Ala Val Gly GlyLeu Ile Asp 3350 3355 3360 Leu Thr Asp Leu Gly Gly Pro Ala His Gly AspAla Gly Ser Trp 3365 3370 3375 Thr Ala Arg Leu Val Leu Leu Arg Arg LeuVal Arg Thr Leu Arg 3380 3385 3390 Gly His Gly Gly Arg Val Leu His ValThr Glu Gly Leu Tyr Gly 3395 3400 3405 Pro Ala Gly Pro Ala Pro Ser LeuAla Gly Ala Arg Met Ala Gly 3410 3415 3420 Phe Val Arg Met Leu Gly AlaGlu Tyr Gly Arg Val Thr Gly Thr 3425 3430 3435 Val Leu Asp Leu Asp ValSer Ala Val Gly Pro Asp Ala Ala Ala 3440 3445 3450 Arg Gln Ile Leu AlaGlu Tyr Thr Gly Pro Tyr Gly Pro Gly Asp 3455 3460 3465 Val Ser Val ArgGly Gly Val Arg His Arg Pro Glu Leu Val Ala 3470 3475 3480 Leu Pro AspAla Gly His Arg Ser Leu Thr Pro Ala Val Asp Arg 3485 3490 3495 Ala TyrLeu Val Thr Gly Gly Thr Arg Gly Ile Gly Ala Arg Val 3500 3505 3510 AlaArg Leu Leu Val Arg Arg Gly Ala Arg Arg Ile Ala Leu Thr 3515 3520 3525Gly Ala Arg Pro Gln Pro Pro Arg Ala Asp Trp Pro Leu Leu Ser 3530 35353540 Pro Gly Thr Pro Glu Ala Glu Thr Ala Ser Leu Val Ala Glu Leu 35453550 3555 Glu Ala Gln Gly Ala Arg Val Leu Val His Ser Gly Pro Leu Ser3560 3565 3570 Glu Arg Glu Arg Thr Asp Arg Phe Leu Arg Glu Val Arg GluVal 3575 3580 3585 Leu Gly Pro Ile Gly Gly Val Val His Cys Ala Gly ArgGly Pro 3590 3595 3600 Val Gly Arg Pro Ser Phe Ile Gly Lys Glu Leu AlaAsp Phe Asp 3605 3610 3615 Pro Val Leu Glu Pro Lys Thr Thr Gly Leu GluVal Leu Asp Glu 3620 3625 3630 Leu Cys Ala Gly Asp Arg Pro Glu Phe PheVal Leu Phe Ser Ser 3635 3640 3645 Leu Ser Ala Val Ala Pro Gly Leu AlaAla Gly Val Leu Asp Tyr 3650 3655 3660 Ala Ala Ala Asn Ala Phe Leu AspCys Tyr Ala Asp His Gln Val 3665 3670 3675 Arg Ser Gly Arg Pro Trp PheArg Ser Val Ala Trp Pro Thr Trp 3680 3685 3690 Ser Glu Ser Gly Met GlyAla Asp Arg Pro Asp Ser Cys Ala Pro 3695 3700 3705 Val Gly Val Gly ProLeu Gly Asp Glu Glu Gly Leu Arg Val Leu 3710 3715 3720 Glu Arg Ile LeuAla Leu Pro Ala Glu Gln Ala Arg Ile Val Pro 3725 3730 3735 Cys Pro ProIle Asp Gly Ile Ala Ala Asp Pro Ala Ala Leu Leu 3740 3745 3750 Gly SerPro Arg Asp Thr Asp Ala Thr Ala Ser Val Gly Ser Thr 3755 3760 3765 ThrSer Ala Gly Ser Thr Pro Met Ala Gly Ser Thr Pro Met Ala 3770 3775 3780Gly Ser Thr Pro Ala Ala Gly Ser Ala Pro Val Pro Thr Thr Thr 3785 37903795 Gly Ala Thr Pro Pro Arg Pro Arg Glu Glu Glu His Thr Val Pro 38003805 3810 Asn Thr Ser Val Thr Gly Pro Pro Trp Leu Ala Pro Leu Phe Ser3815 3820 3825 Glu Leu Leu Ala Ile Pro Glu Asp Ala Leu Asp Pro Thr AlaLeu 3830 3835 3840 Leu Gly Asp Leu Gly Val Glu Ser Val Leu Leu Gly GluIle Leu 3845 3850 3855 Leu Arg Leu Glu Glu Leu Thr Gly Leu Ser Leu AspPro Ala Thr 3860 3865 3870 Leu Leu Asp His Pro Thr Leu Glu Leu Leu GlyArg His Leu Ala 3875 3880 3885 Asp Leu Gly Val Pro Ser Ala Pro Pro AlaPro Ala Ala Thr Thr 3890 3895 3900 Ala Pro Ala Val Ala Pro Val Thr ProVal Thr Pro Thr Ala Pro 3905 3910 3915 Val Ala Val Ala Pro Val Thr ProVal Ala Pro Ser Gly Lys Ile 3920 3925 3930 Ala Val Ile Gly Leu Ser CysArg Phe Pro Gly Ala Glu Asp Ala 3935 3940 3945 Ala Ala Phe Ala Arg AsnLeu Leu Gly Gly Thr Cys Ser Val Thr 3950 3955 3960 Glu Val Pro Pro SerArg Trp Asp Val Gly Glu Leu Tyr Arg Pro 3965 3970 3975 Glu Leu Glu ProGly Arg Ser Thr Ser Lys Trp Gly Gly Phe Leu 3980 3985 3990 Asp Gly IleGlu Asp Phe Asp Pro Glu Trp Phe Gly Met Ser Glu 3995 4000 4005 Asp GluAla Arg Cys Leu Asp Pro Ala Val Arg Leu Phe Leu Glu 4010 4015 4020 GlySer Ala Thr Cys Leu Thr Asp Ala Gly Tyr Gly Ala Arg Glu 4025 4030 4035Leu Ala Gly Arg Asp Val Gly Val Phe Ala Gly Ala Arg Met Ser 4040 40454050 His Tyr Gly Arg Arg Val Gly Glu Arg Arg Gly Leu Val Gly Met 40554060 4065 Gly Ser Asp Gln Asn Phe Ile Ala Ala Arg Ile Ala His His Phe4070 4075 4080 Asp Leu His Gly Pro Asn Leu Val Val Asp Ser Ala Cys SerSer 4085 4090 4095 Ser Leu Val Ala Leu Gln Leu Ala Cys Arg Ser Leu LeuAsp Gly 4100 4105 4110 Glu Ser Glu Leu Ala Leu Ala Gly Gly Val Asp ValLeu Leu Asp 4115 4120 4125 Glu Glu Pro Tyr Leu Asp Phe Ser Ala Ala LysAla Leu Ser Arg 4130 4135 4140 His Gly Arg Cys Ala Thr Phe Asp Glu AspAla Asp Gly Phe Val 4145 4150 4155 Pro Gly Glu Gly Cys Gly Val Val LeuLeu Lys Pro Leu Glu Lys 4160 4165 4170 Ala Leu Arg Asp Gly Asp Arg IleHis Ala Val Ile Asp Ala Val 4175 4180 4185 Ala Val Asn Asn Asp Gly ArgThr Met Gly Leu Thr Thr Pro Asn 4190 4195 4200 Pro Ala Ala Gln Ala LysVal Val Arg Arg Ala Leu Ala Ala Ala 4205 4210 4215 Gly Arg Arg Ala AspGlu Val Gly Leu Ile Glu Ala His Gly Thr 4220 4225 4230 Gly Thr Met IleGly Asp Pro Ile Glu Leu Arg Ala Leu Thr Glu 4235 4240 4245 Val Phe ArgGlu Glu Thr Gly Arg Thr Gly Phe Cys Ala Ile Gly 4250 4255 4260 Ser ValLys Thr Asn Val Gly His Leu Leu Ser Ala Ala Gly Met 4265 4270 4275 AlaGly Leu Ile Lys Ala Val Leu Ala Val Arg Asp Gly Arg Ile 4280 4285 4290Ala Pro Thr Leu Phe Cys Glu Arg Pro Asn Pro Arg Phe Asp Phe 4295 43004305 Ala Ala Ser Pro Phe Tyr Pro Ser Arg Thr Ala His Asp Trp Val 43104315 4320 Pro Glu Pro Gly Arg Val Arg Val Ala Gly Val Ser Ala Phe Gly4325 4330 4335 Leu Gly Gly Thr Asn Ala His Ala Val Val Ser Gln Leu AspPro 4340 4345 4350 Val Leu Ala Ala Ala His Arg Pro Arg Pro Ala Leu ProAla Pro 4355 4360 4365 Asn Phe Ala Arg Arg Arg Leu Trp Leu Glu Ala AlaPro Pro Ser 4370 4375 4380 Ala Pro Gly Pro Arg Pro Ser Val Pro Gly ArgPro Pro Leu Gly 4385 4390 4395 Ala Ser Ile Leu Gly Leu Thr Phe Glu GluPro Leu Ser Glu Pro 4400 4405 4410 Gly Tyr Ala Pro Leu Thr Pro Thr ProThr Gly Ala Asp Thr Ala 4415 4420 4425 Ala Ala Ser Gln Glu Gly Arg LysTrp 4430 4435 46 7349 PRT Streptomyces atroolivaceus 46 Met Asn Val ProSer Ala Pro Gln Pro Gly His Glu Leu Val Cys His 1 5 10 15 Leu Val LeuGlu His Ser Asp Phe Ile Met Gln Asn His Arg Val His 20 25 30 Gly Val SerVal Met Pro Gly Val Thr Phe Leu Asp Ile Val Phe Arg 35 40 45 Ile Leu ArgAsp Arg Gly Phe Asp Thr Ala Arg Ala Glu Leu Arg Asn 50 55 60 Val Leu PheHis Glu Ala Ile Ala Thr Ser Glu Gly Cys Asp Arg Asp 65 70 75 80 Ile ArgIle Thr Val Ser Thr Ser Thr Asp Gly Ser Arg Trp Ile Thr 85 90 95 Ala GluSer Arg Arg Arg Glu Gly Gly Glu Ser Ala Ala Asp Tyr Gln 100 105 110 GluAsn Phe Arg Gly Glu Leu Val Leu His Asp Val Pro Glu Pro Gly 115 120 125Pro Leu Asp Val Asp Arg Leu Arg Thr Thr Ala Arg Arg Val Ala Asp 130 135140 Leu Asp Glu Met Tyr Ala Arg Ala Arg Ala Glu Glu Ile Arg His Gly 145150 155 160 Ser Ala Met Arg Cys Phe Gly Arg Leu Tyr Tyr Gly Asp Gly GluLeu 165 170 175 Leu Ala Glu Leu Gly Leu Asp Gly Glu Ala Ala Ala Leu AspGlu His 180 185 190 Phe His Leu His Pro Ala Lys Met Asp Cys Ala Thr IleAla Ala Phe 195 200 205 Ala Gln Val Pro Pro Pro Asp Gln Asp Pro Phe IlePro Val Phe Ile 210 215 220 Glu Ser Phe Arg Ala Pro Arg Pro Leu Thr GlyVal Ala Tyr Ala His 225 230 235 240 Met Pro Arg Pro Glu Thr Tyr Ala GlnSer Gly Asp Ile Met His Asn 245 250 255 Asp Cys Ala Leu Tyr Asp Ala AspGly Arg Phe Leu Ala Gly Phe Thr 260 265 270 Lys Leu Thr Cys Lys Arg IleArg Asn Pro Glu Leu Ile Thr Arg Leu 275 280 285 Leu Asp Ala Pro Asp ValThr Arg Thr Ala Ala Pro Ala Pro Ala Ala 290 295 300 Val Ser Pro Ser ProVal Val Ala Pro Ala Ser Ser Asp Gly Gly Ala 305 310 315 320 Gly Pro AspAla Val Arg Ala His Leu Arg Glu Leu Val Gly Thr Leu 325 330 335 Leu GlyArg Ala Pro His Ala Ile Arg Thr Asp Ala Gly Phe Tyr Asp 340 345 350 LeuGly Leu Asp Ser Gly His Met Leu Asp Ile Ser Arg Arg Leu Glu 355 360 365Glu Tyr Val Cys Ala Pro Leu Tyr Pro Thr Leu Leu Phe Glu Phe Ser 370 375380 Asp Ile Asp Ser Leu Ala Ala His Leu Tyr Ala Glu Phe Gly Ala Gln 385390 395 400 Val Arg Ser Ala Pro Ala Asn Pro Pro Ala Thr Pro Ala Thr ProGly 405 410 415 Glu Asp Ala Gly Ala Pro Pro Ala Ser Ala Ala Arg Ser ThrAla Arg 420 425 430 Ala Val Ala Pro Ala Leu Gly Cys His Arg Pro Val TrpThr Pro Leu 435 440 445 Pro Ala Asp Pro Gly Ala Phe Ala Ala Asp Gly AlaArg Thr Val Val 450 455 460 Leu Val Gly Ala Asp Ala Ala Thr Ala Ala AlaLeu Arg Asp Ala Ala 465 470 475 480 Ala Pro Ala Arg Val Val Arg Ala GluArg Ala Ser Ala Phe Gln Arg 485 490 495 Leu Ala Ala Asp His Tyr Arg LeuAsp Pro Ala Asp Pro Asp Gln Leu 500 505 510 Ala Ser Leu Thr Ala Ala LeuAla Thr Asp Gly Ile Ser Ala Thr Ala 515 520 525 Tyr Val Arg Cys Ala ArgThr Arg Asp Thr Asp Gly Ala Gly Ser Ala 530 535 540 Leu Pro Asp Ala TyrLeu Glu Ser Trp Ala Leu Ala Val Ala Val Thr 545 550 555 560 Gly Thr ArgPro Thr Gly Pro Val Pro Val Leu Phe Leu His Pro Arg 565 570 575 Asp ProAla Ala Pro Arg Pro His Glu Asp Ala Leu Gly Ala Leu Ala 580 585 590 ArgThr Val Ala Ala Glu Ala Pro Gln Leu Arg Cys Arg Ala Val Gly 595 600 605His Asp Ala Thr Ala Thr Ala Gly Asp Leu Ala Ala Val Ile Ala Ala 610 615620 Glu Ser Thr Asp Leu Ser Ala Glu Ser Glu Val Arg His Thr Gly Gly 625630 635 640 Thr Arg Leu Thr Val Arg His Glu Thr Leu Ala Val Pro Ser GlyAsn 645 650 655 Gly Ala Gly Val Leu Arg Glu Asp Gly Val Tyr Leu Val ThrGly Gly 660 665 670 Gly Gly Ser Leu Ala Ala Leu Leu Val Asp Arg Leu ValThr Arg Gly 675 680 685 Pro Val Arg Leu Val Leu Thr Gly Arg Ser Ala ProGly Pro Glu Leu 690 695 700 Thr Gln Arg Ile Glu Gly Trp Arg Arg Arg GlyAla Glu Val Thr His 705 710 715 720 Val Arg Gly Asp Val Ala His Thr AspAsp Val Leu Ala Ala Val Thr 725 730 735 Cys Ala Arg Glu Thr Tyr Gly ArgIle Asp Gly Val Phe His Cys Ala 740 745 750 Gly Ser Val Asp Asp Gly MetPhe Phe Arg Lys Asp Pro Glu Arg Ser 755 760 765 Ala Ala Val Leu Ala AlaLys Val Ala Gly Thr Arg Asn Leu Asp Glu 770 775 780 Ala Thr Ala Asp AspGly Leu Ala Phe Phe Ala Leu Phe Ser Ser Val 785 790 795 800 Ser Ala SerVal Ala Asn Pro Gly Gln Ala Asp Tyr Ala Tyr Gly Asn 805 810 815 Ala PheMet Glu His Phe Ala Glu Gln Arg Ala Ala Arg Ala Asp Arg 820 825 830 ProGly Val Ser Val Ala Val Gly Trp Pro Leu Trp Ala Asp Gly Gly 835 840 845Met Arg Val Ser Glu Asp Val Leu Arg Arg Ser Ala Asp Thr Ser Gly 850 855860 Leu His Ala Leu Pro Ala Asp Ala Gly Leu Asp Ala Leu Phe Gly Leu 865870 875 880 Leu Ser Gly Ala Ala Pro Arg Ala Val Val Thr Tyr Gly Asp GlnGlu 885 890 895 Arg Ile Ala Glu Leu Leu Pro Ala Pro Arg Pro Ser Ala AlaGln Ser 900 905 910 Gly Arg Thr Gly Ser Pro Asp Ser Pro Asp Ser Pro AspGly Asp Asp 915 920 925 Ile Ala Ile Ile Gly Val Ala Gly Arg Tyr Pro GluAla Glu Asp Leu 930 935 940 Glu Ala Phe Trp Arg Asn Leu Ala Glu Gly ArgAsp Cys Val Gly Glu 945 950 955 960 Val Pro Ala Asp Arg Trp Asp His AlaAla Tyr Tyr Asp Pro Glu Arg 965 970 975 Gly Lys Glu Gly Arg Thr Tyr GlyArg Arg Gly Gly Phe Leu Asp Gly 980 985 990 Val Asp Arg Phe Asp Ala AlaSer Phe Gly Ile Ser Arg Arg Glu Ala 995 1000 1005 Glu Leu Met Asp ProGln Glu Arg Leu Phe Leu Thr Val Gly Arg 1010 1015 1020 Gln Ala Val GluAsn Ala Gly Tyr Arg Pro Glu Glu Leu Ala Arg 1025 1030 1035 Thr Arg ValGly Val Phe Ala Gly Val Met Trp Asn His Tyr Gln 1040 1045 1050 Leu CysThr Asp Gly Ser Ala Glu Pro Val Ala Pro Thr Ala Leu 1055 1060 1065 HisCys Ser Val Ala Asn Arg Leu Ser Tyr Cys Leu Asp Leu Ser 1070 1075 1080Gly Pro Ser Met Ala Val Asp Thr Ala Cys Ser Ser Ser Leu Thr 1085 10901095 Ser Leu His Leu Ala Val Glu Ser Ile Arg Arg Gly Glu Cys Ala 11001105 1110 Leu Ala Val Ala Gly Gly Val Asn Val Ala Ala His Pro Gln Lys1115 1120 1125 Tyr Leu Gln Leu Ala Gln Gly Arg Phe Leu Ser Ser Asp GlyArg 1130 1135 1140 Cys Arg Ala Phe Gly Ala Asp Gly Asp Gly Tyr Val ProGly Glu 1145 1150 1155 Gly Val Gly Ala Val Leu Leu Lys Pro Leu Ala AspAla Leu Ala 1160 1165 1170 Asp Gly Asp His Val His Ala Val Ile Lys GlySer Phe Leu Asn 1175 1180 1185 His Ser Gly Arg Thr Ser Gly Phe Thr ValPro Ser Pro Ala Ala 1190 1195 1200 Gln Ala Thr Leu Ile Ala Asp Ala LeuAsp Arg Ser Gly Val Ala 1205 1210 1215 Ala Asp Ser Val Gly Tyr Ile GluAla His Gly Thr Gly Thr Ala 1220 1225 1230 Leu Gly Asp Pro Ile Glu IleGlu Gly Leu Arg Gln Ala Phe Ala 1235 1240 1245 Asp Ala Gly Leu Ala ProGly Ser Cys Ala Ile Gly Ser Val Lys 1250 1255 1260 Ser Gly Ile Gly HisLeu Glu Ser Ala Ala Gly Ile Ala Ala Val 1265 1270 1275 Thr Lys Val LeuLeu Gln Met Arg His Arg Glu Leu Val Pro Ser 1280 1285 1290 Leu His SerGlu Gln Pro Asn Pro His Ile Asp Phe Ala Ala Thr 1295 1300 1305 Pro PheAla Val Gln Arg Thr Arg Ala Pro Trp Val Pro Arg Pro 1310 1315 1320 GlySer Thr Val Leu Arg Ala Gly Val Ser Ala Phe Gly Ala Gly 1325 1330 1335Gly Ser Asn Ala His Val Leu Leu Glu Ser Ala Pro Pro Ala Pro 1340 13451350 Ala Thr Pro Val Ala Gly Pro Gln Leu Phe Val Phe Ser Ala Lys 13551360 1365 Asp Glu Arg Thr Leu Arg Glu Val Val Arg Arg Gln Leu Arg His1370 1375 1380 Leu Asp Gly Pro Gly Pro Val Gly Ser Ser Ala Asp Glu AlaThr 1385 1390 1395 Ala Leu Leu Thr Gly Glu Val Ala Ala Leu Leu Asp ValPro Val 1400 1405 1410 Asp Ala Val Asp Val Arg Glu Asn Leu Ala Asp LeuGly Val Asp 1415 1420 1425 Arg Leu Ala Leu Ala Glu Leu Gly Arg Arg ValGlu Gly Arg Leu 1430 1435 1440 Pro Ala Gly Val Pro Leu Ser Gly Gln AlaSer Val Thr Glu Leu 1445 1450 1455 Ala Ala Ser Ala Ala Leu Ala Ala ArgPro Asp Ala Leu Pro Leu 1460 1465 1470 Ala Asp Ile Ala His Thr Leu ArgVal Gly Arg Ser Pro Leu Ala 1475 1480 1485 Val Arg Leu Ala Val Val CysGly Glu Pro Glu Glu Leu Arg Arg 1490 1495 1500 Arg Leu Ala Ala Phe LeuAsp Gly Asp Glu Pro Gly Glu Gly Val 1505 1510 1515 Phe Thr Gly Arg AlaAsp Asp Asp Lys Glu Pro Val Arg Leu Glu 1520 1525 1530 Arg Ala Ala GluLeu Phe Arg Leu Gly Arg Leu Ser Glu Leu Ala 1535 1540 1545 Arg Ala TrpAla Asp Gly Ala Ala Val Glu Trp Asp Asp Cys Arg 1550 1555 1560 Ala GlyAsp Gly Val Arg Pro Arg Arg Val Pro Leu Pro Ala His 1565 1570 1575 ProLeu Asp Glu Arg Ser Tyr Trp Ile Gly Gly Trp Arg Ser Ala 1580 1585 1590Gln Glu Thr Gly Glu Ser Arg Glu Pro Val Glu Thr Ala His Ala 1595 16001605 Pro Thr Ala Val Pro Ala Ala Pro Val Asp Gln Ala Asp Leu Pro 16101615 1620 Glu Val Arg Glu Glu Arg Pro Gly Leu Asp Pro Gln Glu Val Leu1625 1630 1635 Trp Ala Val Val Asp Ala Val Arg Thr Arg Leu Tyr Leu GluArg 1640 1645 1650 Asp Glu Val Asp His Arg Leu Ser Phe Asn Glu Met GlyVal Asp 1655 1660 1665 Ser Val Gly Ala Val Glu Ile Val Glu Gln Leu GlyAla Arg Phe 1670 1675 1680 Ala Leu Glu Met Asp Pro Val Thr Leu Phe AspHis Pro Thr Val 1685 1690 1695 Pro Arg Leu Ala Glu His Val Arg Glu LeuHis Arg Gln Ser Pro 1700 1705 1710 Ala Pro Arg Pro Gln Ala Ala Pro AlaPro Ala Gln Pro Ala Ala 1715 1720 1725 Pro Glu Ala Ala Ala Leu Pro AlaPro Ala Pro Ala Pro Ala Pro 1730 1735 1740 Ala Pro Ala Pro Ala Pro AlaSer Lys Pro Glu Pro Ala Ala Ser 1745 1750 1755 Pro Asp Ala Cys Asp AspIle Ala Val Ile Gly Met Ser Gly Arg 1760 1765 1770 Phe Pro Gly Ala GluAsp Leu Asp Ala Phe Trp Glu Asn Ile Ala 1775 1780 1785 Ala Gly Arg AspSer Phe Thr Glu Val Pro Ala Gln Arg Trp Asp 1790 1795 1800 Val Gly ProVal Phe Asp Ala Asp Arg Leu Val Pro Asp Arg Thr 1805 1810 1815 Tyr SerLys Trp Ala Ala Met Leu Pro Glu Val Gly Arg Phe Asp 1820 1825 1830 AlaAla Phe Phe Asn His Ser Pro Leu Glu Ala Glu Val Met Asp 1835 1840 1845Pro Gln Gln Arg Leu Phe Leu Glu Gln Ser Trp Ala Ala Leu Glu 1850 18551860 His Ala Gly Tyr Ala Val Gly Ala Asp Asp Arg Thr Ser Cys Gly 18651870 1875 Val Phe Val Gly Cys Ala Pro Gly Asp Tyr Ser Thr Leu Leu Thr1880 1885 1890 Glu Ala Gly Arg Ala Asp Thr Gly His Ala Phe Leu Gly ThrThr 1895 1900 1905 Ser Ser Leu Leu Pro Ala Arg Ile Gly Tyr Phe Leu AsnLeu Asp 1910 1915 1920 Gly Pro Thr Met Ala Val Asp Thr Ala Cys Ser SerSer Leu Val 1925 1930 1935 Ala Val His Leu Ala Ala Asp Ser Ile Arg ArgGly Glu Cys Ala 1940 1945 1950 Met Ala Leu Ala Gly Gly Val Ala Leu MetVal Thr Pro Gln Leu 1955 1960 1965 His Val Arg Ala Ser Lys Val Gly MetLeu Ser Pro Arg Gly Thr 1970 1975 1980 Cys Val Pro Phe Asp Ala Ser AlaAsp Gly Thr Val Leu Gly Glu 1985 1990 1995 Gly Val Gly Ala Val Val LeuLys Arg Leu Asp Arg Ala Val Ala 2000 2005 2010 Asp Gly Asp His Ile HisGly Val Ile Lys Ala Thr Gly Val Asn 2015 2020 2025 Gly Asp Gly Arg ThrAsn Gly Ile Thr Ala Pro Ser Ala Leu Ser 2030 2035 2040 Gln Ala Ala LeuIle Ala Asp Val His Arg Arg Ala Gly Val Gly 2045 2050 2055 Ala Asp AspIle Gly Tyr Val Glu Ala His Gly Thr Gly Thr Ala 2060 2065 2070 Leu GlyAsp Pro Ile Glu Val Arg Ala Leu Thr Glu Val Phe Arg 2075 2080 2085 ArgSer Thr Asp Arg Ser Gly Tyr Cys Gly Ile Gly Thr Val Lys 2090 2095 2100Ala Asn Ile Gly His Thr Thr Met Ala Ala Gly Ile Ala Gly Leu 2105 21102115 Leu Lys Thr Leu Leu Ala Leu Arg His Ser Glu Leu Pro Pro Ala 21202125 2130 Pro Ala Phe Asp Thr Pro Asn Pro Lys Thr Glu Leu Asp Ser Ser2135 2140 2145 Pro Phe Phe Val Val Arg Asp Arg Gln Glu Trp Glu Pro GlyPro 2150 2155 2160 Gly Gly Gln Arg Ile Ala Thr Val Ser Ser Phe Gly PheSer Gly 2165 2170 2175 Thr Asn Ala His Ala Val Leu Ala Gln Ala Pro GluPro Gln Ala 2180 2185 2190 Arg Pro Glu Glu Pro Asp Gln Glu Arg Leu PheAla Val Ser Ala 2195 2200 2205 Arg Asp Gly Ala Ala Leu Asp Arg Leu LeuLeu Arg Leu Ala Asp 2210 2215 2220 Ser Asp Leu Asp Gly Val Thr Pro AlaAsp Leu Ala Phe Thr Leu 2225 2230 2235 Gly Val Gly Arg Ala His Leu ProVal Arg Ala Ala Val Ile Ala 2240 2245 2250 Arg Asn Val Pro Glu Leu ArgArg Arg Leu Arg Leu Leu Gln Ser 2255 2260 2265 Gly Ala Gln Ala Pro GlyCys Phe Arg Thr Gly Gln Gly Ala Ala 2270 2275 2280 Ala Gly Asp Leu AspGlu Gln Thr Arg Ala Glu Leu Ala Gly Arg 2285 2290 2295 Ala Arg Ser GlySer Pro Ala Glu Arg Val Ala Ala Leu Glu Arg 2300 2305 2310 Leu Ala AlaAla Tyr Ala Ala Gly Gln Asp Leu Asp Trp Gln Ser 2315 2320 2325 Leu SerTyr Gly Asp Arg Pro Arg Arg Val Pro Leu Pro Thr Tyr 2330 2335 2340 ProPhe Gly Gly Asp Arg His Trp Ile Thr Leu Pro Asp Thr Asp 2345 2350 2355Arg Thr Ala Val Pro Ala Thr Ala Pro Ala Thr Ser Arg Val Asp 2360 23652370 Leu Pro Gly Gln Ser Pro Gln Ser Thr Pro His Pro Leu Leu Gly 23752380 2385 Ala Val Ser Gly Ala Pro Gly Asp Pro Asp Gly Ala Arg Phe Pro2390 2395 2400 Val Pro Val Pro Ala Ala His Trp Val Leu Asp His His ArgIle 2405 2410 2415 Gly Asp Arg Pro Val Leu Pro Gly Ala Ala Gly Leu AspLeu Ala 2420 2425 2430 Val Ala Ala Ala Arg Arg Cys Gly Leu Arg Gly ThrVal Arg Leu 2435 2440 2445 His Gly Val Gln Trp Leu Arg Leu Ile Asp AlaGlu Ala Ala Gly 2450 2455 2460 Thr Leu Arg Leu Thr Leu Thr Ser Asp GlyGlu Gly Tyr Arg Phe 2465 2470 2475 Ala Leu Ser Thr Gly Asp Asp Gly ThrVal Cys Ser Arg Gly Ser 2480 2485 2490 Leu Thr Val Arg Pro Asp Ala GlnAsp Ala Ala Ala Pro Ala Ala 2495 2500 2505 Ser Ser Glu Thr Leu Asp ValAla Glu Ile Ala Ala Arg Cys Pro 2510 2515 2520 Tyr Glu Val Pro Ala GluArg Phe Tyr Asp Asp Phe Arg Ser Gly 2525 2530 2535 Gly Ile Ala Tyr GlyPro Ser Phe Arg Val Leu Glu Lys Ile Thr 2540 2545 2550 Phe Gly Asp AspGlu Val Leu Gly Thr Leu Arg Ala Thr Pro Asp 2555 2560 2565 Ser Gly GlyPhe Ala Leu His Pro Ala Leu Leu Asp Gly Ala Gln 2570 2575 2580 Gln ThrIle Ala Ala Leu Glu Gly Gly Asn Asp Ala Thr Leu Val 2585 2590 2595 ProPhe Ser Val Glu Thr Val Glu Val Val Asp Ala Thr Ala Val 2600 2605 2610Pro Ala Phe Ala His Val Val Arg Ala Gly Lys His Arg Tyr Thr 2615 26202625 Val Arg Leu Ala Asp Arg Ser Gly Arg Val Cys Val Arg Tyr Glu 26302635 2640 Gly Leu Ala Leu Arg Ala Gln His Asn Pro Val Asp Ser Met Met2645 2650 2655 Tyr Arg Pro Val Trp Arg Pro Ala Pro Leu Pro Gln Pro GlyAsn 2660 2665 2670 Ala Pro Ala Gly Gly Arg Thr Val Val Val His Thr AlaAsp Ser 2675 2680 2685 Thr Thr Leu Ala Ala Ala Leu Ala Ala Arg Thr GlyAla Gly Leu 2690 2695 2700 Val Ala Leu Ser Gly Ala Gln Asp Ala Ala ProAsp Pro Tyr Ala 2705 2710 2715 Val Leu Glu Gln Pro Leu Glu Thr Val TyrPhe Val Ala Arg Thr 2720 2725 2730 Gly Asp Ala Glu Gly Pro Ala Glu AlaAsp Arg Thr Ala Leu Asp 2735 2740 2745 Leu Phe Arg Leu Val Lys Arg MetLeu Ala Val Gly Arg Ala Arg 2750 2755 2760 Asp Arg Ile Ala Leu Arg IleVal Leu Ala Gly Ala Val Pro Ala 2765 2770 2775 Asp Pro Glu Asp Met ThrGlu Thr Val Arg Pro His Ala Ala Gly 2780 2785 2790 Val Leu Gly Leu AlaArg Ala Ile Glu Ser Glu Cys Pro Arg Trp 2795 2800 2805 Ser Val Ala CysVal Asp Val Gly Ala Asp Gly Gly Thr Val Gly 2810 2815 2820 Ala Glu ArgAla Ala Glu Arg Ile Val Ala Glu Pro Gly Thr Glu 2825 2830 2835 Pro LeuVal Leu Leu Arg Gly Glu Glu Arg Leu Glu Arg Val Phe 2840 2845 2850 GluPro Leu Arg Pro Ala Ala Pro Arg Gly Thr Glu Pro Phe Arg 2855 2860 2865Glu Gly Gly Val Tyr Val Ile Val Gly Gly Ala Gly Gly Ile Gly 2870 28752880 Phe Ala Leu Ser Arg Leu Leu Ala Arg Ile Ala Arg Ala Arg Leu 28852890 2895 Val Trp Ile Gly Arg Ser Pro Glu Gly Pro Glu His Arg Ala Lys2900 2905 2910 Ala Glu Glu Ile Ala Ala Leu Gly Gly Gln Val Leu Tyr ValGln 2915 2920 2925 Ala Asp Val Ala Asp Glu Ala Ala Leu Arg Arg Gly LeuAla Ser 2930 2935 2940 Val His Thr Arg Phe Gly Gln Val Asp Gly Ala ValHis Ala Ala 2945 2950 2955 Leu Asp Leu Arg Asp Arg Thr Ile Ala Leu MetAsp Glu Glu Asp 2960 2965 2970 Phe Leu Ala Gly Leu Ala Pro Lys Val AlaGly Val Thr Ala Phe 2975 2980 2985 Ala Arg Val Phe Gly Ala Glu Pro LeuAsp Phe Met Leu Val Phe 2990 2995 3000 Ser Ser Ala Val Ser Phe Val GluAla Gly Gly Gln Ala Asn Tyr 3005 3010 3015 Ala Ala Ala Ser Thr Phe GluAsp Ala Tyr Val Gln Trp Leu Asp 3020 3025 3030 Arg Arg His Asp Tyr ProVal Ser Val Val Asn Trp Gly Phe Trp 3035 3040 3045 Gly Ser Val Gly AlaVal Ala Asp Asp Arg Met Arg Ala Ala Phe 3050 3055 3060 Ala Arg Leu GlyVal Gly Ser Val Glu Pro Ala Glu Gly Met Ala 3065 3070 3075 Val Leu ArgArg Ile Ile Ala Gly Arg Leu Pro Gln Thr Leu Ala 3080 3085 3090 Met LysAla Asp Arg Ala Ala Leu Pro Ala Met Gly Ile Arg Val 3095 3100 3105 AlaGly Thr Pro Glu Gly Glu Gly Pro Ala Arg Thr Ser Pro Pro 3110 3115 3120Ala Thr Ser Ala Ala Pro Gly Pro Gly Thr Thr Thr Pro Ala Pro 3125 31303135 Ser Pro Ala Gln Val Ala Val Pro Asp Pro Val Ala Ala Gly Ala 31403145 3150 Glu Gly Leu Ser Ala Glu Ala Val Arg Ser Tyr Val Ala Gly Val3155 3160 3165 Phe Ala Glu Val Leu Lys Tyr Glu Ser Ala Ala Leu Asp ProGlu 3170 3175 3180 Ala Thr Phe Glu Thr Phe Gly Val Asp Ser Leu Val SerLeu Asn 3185 3190 3195 Ile Val Asp Arg Phe Glu Gln Asp Leu Gly Asp LeuPro Gln Thr 3200 3205 3210 Leu Leu Phe Glu Tyr Met Thr Ile Asp Gln ValAla Gly Tyr Phe 3215 3220 3225 Arg Gln Glu His Gly Glu Gln Leu Thr ArgVal Ile Ala Pro Ala 3230 3235 3240 Pro Thr Gln Asp Gln Gly Gln Gly AlaVal Ala Ala Ser Pro Ala 3245 3250 3255 Ala Glu Val Val Ser Gly Ala AlaVal Gln Gly Gly Asp Glu Glu 3260 3265 3270 Val Phe Ala Arg Ala Gln GluAla Phe Ala Ala Val Glu Ala Phe 3275 3280 3285 Ser Arg Asp Leu Leu ArgArg Thr Phe Pro Arg Leu Asp Gly Val 3290 3295 3300 Pro Arg Pro Gly GluArg Ile Thr Ala Asp Glu Leu Ala Ser Arg 3305 3310 3315 Leu Gly Val ValArg Arg His Arg Arg Leu Phe Asp Ala Ala Leu 3320 3325 3330 Ser Ile LeuArg Ser Cys Gly Ala Val Thr Gly Asp Ala Asp Thr 3335 3340 3345 Leu ThrPhe Ala Glu Pro Ser Ala Ala Pro Gly Ala Arg Val Glu 3350 3355 3360 GlyAla Glu Val Ala Ala Leu Tyr Pro Glu Met Ser Gly His Val 3365 3370 3375Thr Leu Leu Glu Arg Thr Leu Gly Ala Leu Gly Glu Val Leu Ala 3380 33853390 Gly Arg Arg Asn Pro Met Asp Val Leu Phe Pro Lys Gly Ser Val 33953400 3405 Ala Leu Val Glu Pro Ile Tyr Lys Gly Gln Pro Ile Ala Asp His3410 3415 3420 Tyr Asn Arg Leu Leu Ala Asp Glu Val Ala Asp Ala Ala ArgArg 3425 3430 3435 Val Arg Ala Gln Glu Gly Arg Pro Val Arg Val Leu GluIle Gly 3440 3445 3450 Ala Gly Thr Gly Ala Ser Ser Arg Thr Val Leu AlaAla Leu Ala 3455 3460 3465 Ala Ala Asp Ala Gly Ala His Tyr Cys Tyr ThrAsp Ile Ser Pro 3470 3475 3480 Ala Phe Leu Arg His Gly Glu Arg Glu PheGly Pro Thr Tyr Pro 3485 3490 3495 Gln Leu Ala Phe His Thr Leu Asp IleSer Arg Asp Pro Val Glu 3500 3505 3510 Gln Gly Ile Glu Ala Ala Ser TyrAsp Val Ile Leu Gly Thr His 3515 3520 3525 Val Leu His Ala Thr Pro AspMet Glu Arg Thr Leu Arg Asn Ile 3530 3535 3540 Arg Thr Leu Leu Arg ProGly Gly Leu Val Leu Val Asn Glu Ile 3545 3550 3555 Thr Arg Phe Ser GluPhe Leu Thr Leu Thr Phe Gly Leu Thr Thr 3560 3565 3570 Gly Trp Trp MetTyr Glu Asp Ala Gln Cys Arg Leu Pro His Ser 3575 3580 3585 Pro Leu LeuAla Pro Val Gln Trp Arg Gln Ser Ala Ala Ala Ala 3590 3595 3600 Gly LeuArg Thr Val Arg Thr Gly Gly Leu Pro Gly Val Pro Ala 3605 3610 3615 AspGlu Leu Glu Gln Ser Leu Val Val Ala Glu Arg Pro Val Glu 3620 3625 3630Asp Ser Gly Asp Ala Ser Pro Asp Gly Ala Ala Asp Glu Gln Ser 3635 36403645 Pro Glu Ser Val Arg Ser Tyr Val Thr Gly Val Phe Ala Glu Val 36503655 3660 Leu Lys Tyr Arg Ala Glu Asp Leu Asp Pro Ala Val Thr Leu Glu3665 3670 3675 Asn Phe Gly Val Asp Ser Leu Val Ser Leu Asn Ile Val AspArg 3680 3685 3690 Leu Glu Gln Asp Leu Gly Asp Leu Pro Gln Thr Leu LeuPhe Glu 3695 3700 3705 Tyr Thr Ser Ile Asp Ser Ile Ala Glu Tyr Leu SerAla Glu His 3710 3715 3720 Gly Glu Arg Leu Ala Arg Val Leu Gly Gly AlaPro Ala Ala Ala 3725 3730 3735 Gln Ala Gln Pro Ser Ala Pro Val Pro AlaPro Val Ser Val Asp 3740 3745 3750 Val Pro Ala Pro Ile Pro Ala Pro IlePro Ala Pro Val Ser Val 3755 3760 3765 Pro Val Asp Val Gln Glu Pro GluSer Glu Pro Glu Pro Ala Ala 3770 3775 3780 Ala Val Arg Thr Pro Ala GlyAsp Asp Pro Ala Asp Pro Leu Asp 3785 3790 3795 Ile Ala Val Ile Gly ValVal Gly Arg Tyr Pro Gln Ser Pro Asp 3800 3805 3810 Leu Glu Ala Phe TrpArg Asn Leu Ser Glu Gly Arg Ser Cys Ile 3815 3820 3825 Thr Glu Ile ProSer Glu Arg Trp Asp Trp Arg Arg Asn Phe Asp 3830 3835 3840 Pro Asp LysSer Arg Lys His Arg Ser Tyr Ser Arg Trp Gly Gly 3845 3850 3855 Phe LeuGlu Asp Ile Glu Met Phe Asp Ala Pro Leu Phe Gly Ile 3860 3865 3870 LeuPro Arg Asp Ala Ala Asp Ile Asp Pro Gln Glu Arg Leu Phe 3875 3880 3885Leu Glu Ser Cys Trp Glu Leu Leu Glu Thr Ala Gly Tyr Leu Gly 3890 38953900 Thr Tyr Thr His Glu Pro Gln Thr Gly Val Phe Ala Gly Leu Met 39053910 3915 Tyr Gly Glu Tyr Gly Leu Leu Ala Ala Ala Thr Asp Trp Pro Glu3920 3925 3930 Gly Arg Tyr Ala Thr Gly His Ser Ala Tyr Trp Ser Met AlaAsn 3935 3940 3945 Arg Val Ser Tyr Thr Phe Asp Leu Gln Gly Pro Ser LeuAla Val 3950 3955 3960 Asp Ser Ala Cys Ser Ser Ala Leu Ser Ala Ile GlnLeu Ala Cys 3965 3970 3975 Glu Ser Leu Arg Arg Gly Glu Ser Arg Met AlaIle Ala Gly Gly 3980 3985 3990 Thr Asn Leu Ile Leu His Pro Ala His PheAla Ala Leu Cys Ala 3995 4000 4005 Arg Asn Met Leu Ser Ala Ala Asp AlaCys Arg Val Phe Asp Asp 4010 4015 4020 Gly Ala Asp Gly Phe Val Pro GlyGlu Gly Ala Gly Ala Val Leu 4025 4030 4035 Leu Lys Pro Leu Ala Gln AlaGlu Ala Asp Gly Asp Thr Ile Trp 4040 4045 4050 Gly Val Val Lys Gly AlaPhe Ser Asn Ala Gly Gly Lys Val Ser 4055 4060 4065 Gly Tyr Thr Val ProAsn Pro Asn Ala Gln Ala Arg Leu Val Glu 4070 4075 4080 Arg Thr Leu ArgArg Ser Gly Val His Pro Arg Thr Val Ser Tyr 4085 4090 4095 Val Glu AlaHis Gly Thr Gly Thr Ala Leu Gly Asp Pro Ile Glu 4100 4105 4110 Leu GlyGly Leu Thr Lys Ala Phe Arg Ala Ala Gly Ala Thr Gly 4115 4120 4125 AspGly Tyr Cys Ala Val Gly Ser Val Lys Ser Asn Ile Gly His 4130 4135 4140Leu Glu Gly Ala Ala Gly Ile Ala Ala Val Thr Lys Val Leu Leu 4145 41504155 Gln Leu Lys His Arg Ala Leu Ala Pro Thr Ile His Leu Asp Arg 41604165 4170 Leu Asn Pro Lys Ile Asp Phe Ala Gly Ser Pro Phe Gly Pro Gln4175 4180 4185 Arg Thr Ala Glu Pro Trp Asp Arg Pro Val Ala Gly Val AspGly 4190 4195 4200 Ala Glu Arg Ser Trp Pro Arg Arg Ala Gly Ile Ser SerPhe Gly 4205 4210 4215 Ala Gly Gly Ala Asn Val His Met Ile Leu Glu GluTyr Thr Gly 4220 4225 4230 Gln Asp Pro Arg Asp Ala Glu Asp Thr Met GlyAla Ala Gly Ala 4235 4240 4245 Glu Glu Pro Glu Leu Phe Val Leu Ser AlaLeu Asp Arg Glu Thr 4250 4255 4260 Leu Ala Arg His Ala Gly Arg Val AlaAsp Phe Val Ala Gly Pro 4265 4270 4275 Glu Gly Ala Arg Val Arg Leu AlaAsp Leu Ala His Thr Ser Arg 4280 4285 4290 Val Gly Arg Arg Glu Leu ProGlu Arg Leu Ala Val Thr Ala Ala 4295 4300 4305 Ser His Ala Gln Leu AlaAla Arg Leu Arg Glu Phe Ala Ala Thr 4310 4315 4320 Gly Val Ala Gly GluGly Val Ser Thr Gly Thr Ala Arg Lys Gly 4325 4330 4335 Gly Ala Gly SerGly Leu Gly Ala Gln Glu Leu Thr Ala Ala Leu 4340 4345 4350 Ala Gly ArgArg Trp Ala Asp Ala Val Glu His Trp Thr Leu Gly 4355 4360 4365 Gly ArgVal Asp Trp Arg Thr Ala Asp Ala Gly Arg Leu Val Arg 4370 4375 4380 LysVal Ala Phe Pro Thr Tyr Pro Phe Asn Arg Ser Arg His Trp 4385 4390 4395Ile Thr Ser Asp Pro His His Leu Ala Pro Glu Leu Ala Ser Gly 4400 44054410 Gly Arg Pro Ala Glu Glu Pro Leu Ser Gly Ser Glu Pro Ala Ala 44154420 4425 Pro Glu Pro Ala Ala Thr Arg Leu Pro Ser Pro Glu Pro Ala Gly4430 4435 4440 Pro Glu Pro Val Ala Ala Glu Arg Pro Val Ala Val Pro LeuPro 4445 4450 4455 Val Pro Asp Leu Val Gln Pro Val Glu Asp Asn Asp ArgAsp Glu 4460 4465 4470 Gly Gly Pro Ala Ala Asp Val Thr Ala Glu Arg GlyAla Thr His 4475 4480 4485 Pro Ala Leu Ala Leu Ala Ala His His Arg IleSer Gly Ala Arg 4490 4495 4500 Trp Val Pro Gly Val Ser Leu Leu Glu ValAla Arg Glu Ala Cys 4505 4510 4515 Pro Gly Ala Asp Gly Ser Ala Gly AlaGlu Phe Pro Phe Arg Leu 4520 4525 4530 Gly Ser Val Arg Trp Leu Thr ProVal Asp Leu Asp Ala Gly Thr 4535 4540 4545 Asp Arg Val Thr Val Gly PheThr Asp Asp Gly Thr Gly Thr Ala 4550 4555 4560 Phe Arg Ile Thr Ala GlyGly Pro Ser Gly Thr Thr Val Ala Arg 4565 4570 4575 Gly Thr Leu Thr GluGly Pro Cys Ala Asp Asp Ala Gly Ile Asp 4580 4585 4590 Leu Ala Ala ValArg Asp Arg Ser Thr Gly Arg Pro Asp Pro Cys 4595 4600 4605 Glu Phe TyrAsp Ala Phe Ala Arg Ala Gly Leu Glu Tyr Gly Arg 4610 4615 4620 Pro LeuArg Thr Val Ala Glu Leu Trp Thr Gly Thr Gly Glu Cys 4625 4630 4635 LeuAla Arg Leu Ser Leu Ser Arg Leu Met Arg Arg Val Gly Pro 4640 4645 4650Ala His Ala Leu His Pro Ala Leu Leu Asp Gly Ala Leu Gln Ala 4655 46604665 Val Ser Ser Leu Cys Ala Ala Asp Glu Ser Tyr Leu Pro Val Gly 46704675 4680 Met Gly Glu Ile Arg Ala His Gly Ala Leu Pro Ala Asp Ser Trp4685 4690 4695 Ala Tyr Val Arg Glu Val Thr Ala Glu Gly Gly Thr Gly GlyArg 4700 4705 4710 Arg Arg Phe Asp Ile Arg Leu Thr Asp Gly Thr Gly ArgVal Leu 4715 4720 4725 Gln Ala Val Asp Gly Leu Glu Ile Ala Pro Arg AlaGly Gly Gly 4730 4735 4740 Ala Pro Glu Thr Arg Thr Arg Tyr Leu Ala ProVal Trp Thr Pro 4745 4750 4755 Ala Pro Leu Pro Ala Gly Glu Gln Ala ProGly Thr Val Leu Leu 4760 4765 4770 Cys Gly Ala Glu Thr Pro Gln Arg GlnGlu Leu Ala Gly Gln Leu 4775 4780 4785 Thr Ala Ala Gly Ser Thr Val ValCys Ala Ser Asp Gly Pro Ala 4790 4795 4800 Phe Ala Arg Ser Asp Ala GlyPro His Ala Ala Tyr Thr Leu Arg 4805 4810 4815 Gly Thr Asp Pro Ala GluHis Thr Arg Leu Ile Glu Asp Leu Ala 4820 4825 4830 Gly Arg Gly Leu LeuPro Asp Ala Val Val His Leu Ala Asp Glu 4835 4840 4845 Ala Glu Ala AspIle His Arg Gln Asp Val Asp Ser Gly Glu Ser 4850 4855 4860 Val Leu SerPhe Val Leu Trp Ser Ala Val Ala Val Leu Gly Arg 4865 4870 4875 Pro GluArg Pro Gly Leu Arg Val Val Phe Ala His Arg Ala Gly 4880 4885 4890 AlaGln Gly Ala Glu Pro Leu His Ile Ala Val Gly Ala Thr Leu 4895 4900 4905Arg Thr Leu Gly Leu Glu His Ser Arg Phe Ser Gly Val Arg Val 4910 49154920 Glu Leu Pro Ala Gly Gly Ala Gly Thr Asp Glu Val Val Ala Glu 49254930 4935 Leu Arg Ala Ala Arg Pro Gly Ala Val Glu Val Ser Tyr Ala Ala4940 4945 4950 Asp Gly Gly Arg Ser Glu Lys Asn Leu Glu Pro Phe Glu ProPro 4955 4960 4965 Ala Ala Gly Leu Cys Asp Phe Ala Pro Arg Ala Gly GlyThr Tyr 4970 4975 4980 Leu Ile Thr Gly Gly Ala Gly Ala Leu Gly Leu HisPhe Ala Ala 4985 4990 4995 Tyr Leu Ala Gly Arg Val Pro Asp Asp Ala ThrIle Val Leu Ala 5000 5005 5010 Gly Arg Ser Gln Pro Gly Gly Asp Ala AlaArg Arg Ile Ala Glu 5015 5020 5025 Leu Asn Gly Asp Gly Ala Arg Val LeuHis Arg Pro Ala Asp Leu 5030 5035 5040 Arg Asp Ala Ala Ser Val Glu ArgLeu Val Ala Ser Val Ser Ala 5045 5050 5055 Glu Phe Gly Pro Leu Arg GlyVal Leu His Ser Ala Gly Val Thr 5060 5065 5070 Arg Asp Ala Arg Ala ValLys Lys Thr Ala Arg Glu Leu Asp Glu 5075 5080 5085 Val Leu Ala Pro LysVal Ser Gly Thr Val His Leu Asp Ala Ala 5090 5095 5100 Thr Ala Asp GlnPro Leu Asp Phe Phe Val Leu Phe Ser Ser Met 5105 5110 5115 Ser Gly GluSer Gly Asn Leu Gly Gln Ala Asp Tyr Ala Tyr Ala 5120 5125 5130 Asn AlaPhe Leu Asn Ala Phe Ala Ala Arg Arg Asp Gly Met Arg 5135 5140 5145 AlaAla Gly Leu Arg Ser Gly Arg Ser Leu Ser Ile Gly Trp Pro 5150 5155 5160Leu Trp Ala Asp Gly Gly Met Thr Val Asp Asp Ala Thr Arg Glu 5165 51705175 Met Phe Ala Arg Thr Trp Asp Met Val Pro Leu Ser Thr His Ala 51805185 5190 Gly Leu Glu Ala Phe Val Arg Gly Leu Ala Cys Glu Leu Pro Ala5195 5200 5205 Leu Cys Val Val Glu Ser Leu Val Arg Arg Glu Ala Ala AlaPro 5210 5215 5220 Ala Thr Ala Pro Arg Pro Ala Ala Gly Arg Gln Glu LeuPro Ala 5225 5230 5235 Met Asp Val Val Ala Leu Arg Glu Leu Val Glu HisGlu Ala Arg 5240 5245 5250 Val Leu Ala Ser Gly Phe Leu Leu Val Asp ProSer Glu Val Asp 5255 5260 5265 Val Ala Ala Glu Leu Leu Glu Leu Gly PheAsp Ser Ile Thr Leu 5270 5275 5280 Thr Glu Leu Val Asn Lys Val Asn GluArg Phe Gly Leu Asp Leu 5285 5290 5295 Leu Pro Thr Val Leu Phe Glu CysPro Asp Leu Val Ser Phe Ala 5300 5305 5310 Glu Tyr Leu Thr Gln His HisAla Ala Glu Leu Ala Gln His Tyr 5315 5320 5325 Ala Pro Asp Ser Pro AlaGlu Arg Thr Glu Val Arg Asp Glu Pro 5330 5335 5340 Ala Val Ser Asp GlyThr Pro Ala Pro Ala Pro Ala Ser Pro Val 5345 5350 5355 Val Pro Ala SerPro Ala Pro Ser Val Pro Thr Val Thr Ala Val 5360 5365 5370 Thr Thr SerPro Ala Pro Ala Pro Ala Pro Ala Pro Ala Ala Arg 5375 5380 5385 Ala GluIle Ala Val Ile Gly Ile Ala Gly Val Phe Pro Gly Ser 5390 5395 5400 AlaAsp Thr Asp Glu Phe Trp Glu His Leu Ala Gly Gly Val Asp 5405 5410 5415Leu Val Arg Pro Val Pro Lys Asp Arg Thr Ala Ile Arg Ala Asn 5420 54255430 Pro Ala Thr Arg Glu Leu Arg Gly Gly Phe Leu Asp Ser Val Asp 54355440 5445 Thr Phe Asp Ala Arg Leu Phe Gly Ile Ser Pro Asn Glu Ala Ala5450 5455 5460 Leu Met Asp Pro Gln Gln Arg Leu Phe Leu Gln Thr Ala TrpArg 5465 5470 5475 Val Phe Glu Asp Ala Gly Tyr Arg Pro Ala Asp Leu AlaGly Ala 5480 5485 5490 Pro Cys Gly Leu Phe Val Gly Val Ala Thr His AspTyr Asp Asp 5495 5500 5505 Leu Leu Lys Glu Asn Gly Val Ala Val Gln AlaHis Thr Ala Thr 5510 5515 5520 Gly Ile Ala His Ser Val Leu Ala Asn ArgVal Ser Tyr Leu Phe 5525 5530 5535 Asp Leu Asn Gly Pro Ser Glu Ala ValAsp Thr Ala Cys Ser Ser 5540 5545 5550 Ser Leu Val Ala Ile His Arg AlaLeu Arg Ala Ile Gln Asp Gly 5555 5560 5565 Glu Cys Glu Leu Ala Val AlaGly Gly Val Asn Val Ile Leu Thr 5570 5575 5580 Pro Gly Leu Leu Glu SerPhe Thr Gln Ser Gly Met Leu Ser Pro 5585 5590 5595 Asp Gly Arg Cys LysThr Phe Asp Ala Asp Ala Asp Gly Tyr Val 5600 5605 5610 Arg Gly Glu GlyVal Gly Ala Val Leu Leu Lys Pro Leu Ala Arg 5615 5620 5625 Ala Glu AlaAsp Gly Asp His Ile Tyr Ala Val Val Lys Gly Thr 5630 5635 5640 Ala ValAsn His Gly Gly Arg Ser Asn Ser Leu Thr Ala Pro Asn 5645 5650 5655 ProGlu Ser Gln Ala Arg Val Val Ala Ala Ala Val Arg Glu Ala 5660 5665 5670Gly Val Glu Pro Asp Thr Ile Thr Tyr Ile Glu Ala His Gly Thr 5675 56805685 Gly Thr Arg Leu Gly Asp Pro Ile Glu Ile Glu Gly Leu Lys Lys 56905695 5700 Ala Phe Thr Thr Leu His Glu Glu Arg Gly Glu Ala Val Pro Asp5705 5710 5715 Thr Gly Arg Ile Ala Ile Gly Ala Val Lys Thr Asn Ile GlyHis 5720 5725 5730 Leu Glu Thr Ala Ser Gly Ile Ala Gly Val Leu Lys ValVal Gln 5735 5740 5745 Ser Met Arg His Arg Val Leu Pro Ala Ser Leu HisLeu Arg Arg 5750 5755 5760 Leu Ser Pro Tyr Leu Arg Leu Asp Gly Thr ProPhe Thr Val Asn 5765 5770 5775 Asp Arg His Arg Pro Trp Glu Pro Ala LeuThr Pro Asp Gly Arg 5780 5785 5790 Gln Val Leu Arg Ala Gly Val Ser SerPhe Gly Phe Gly Gly Ser 5795 5800 5805 Asn Ala His Val Val Leu Glu AlaTyr Pro Ala Arg Thr Ala Pro 5810 5815 5820 Ala Val Gln Asp Phe Ala ProHis Thr Val Pro Leu Ser Ala Gly 5825 5830 5835 Asp Pro Asp Asp Leu ArgGly Tyr Ala Ala Arg Leu Ala Arg His 5840 5845 5850 Leu Ala Arg Thr ProGlu Ala Asp Leu Ala Arg Val Ala Tyr Thr 5855 5860 5865 Leu Gln Thr GlyArg Thr Gly His Arg His Arg Phe Ala Val Arg 5870 5875 5880 Val Arg AspArg Asp Glu Leu Ile Gly Ala Leu Glu Ala Phe Ala 5885 5890 5895 Ala GlyGlu Leu Pro Asp His Ala Ala Thr Gly Thr Ala Arg Arg 5900 5905 5910 AspAla Pro Ser Val Gln Ser Asp Glu Asp Pro Ala Leu Leu Arg 5915 5920 5925Lys Ser Trp Cys Glu Gly Ala Asp Val Pro Trp His Thr Trp Trp 5930 59355940 Pro Lys Thr Pro Gly Arg Val Pro Leu Pro Thr Ala Pro Phe Ala 59455950 5955 Arg Thr Arg His Trp Phe Pro Thr Pro Asp Ala Glu Pro Thr Ala5960 5965 5970 Pro Ala Val Pro Ala Arg Lys Glu Ala Thr Met Thr Ala ArgSer 5975 5980 5985 Glu Ala Gln Pro Thr Arg Arg Gly Pro Lys Ile Arg LeuAla Ala 5990 5995 6000 Pro Arg Ser Ala Gly Gly Ala Ser Ala Thr Ala ProThr Arg Glu 6005 6010 6015 Pro Val Pro Ala Pro Pro Ala Pro Ala Pro AlaPro Ala Thr Thr 6020 6025 6030 Ala Pro Gln Pro Val Arg Ala Asp Val ProPro Pro Ala Leu Pro 6035 6040 6045 Glu Ala Ala Ala Ala Ser Thr Val ValGly Leu Leu Arg Gly Glu 6050 6055 6060 Leu Ser Lys Ile Leu Gly Met ProSer Glu Glu Ile Glu Asn Asp 6065 6070 6075 Ala Pro Phe Gly Glu Leu GlyLeu Asp Ser Ile Tyr Arg Met Asp 6080 6085 6090 Leu Val Arg Thr Leu AsnGlu Ala Phe Gly Leu Asp Leu Lys Ala 6095 6100 6105 Thr Glu Leu Tyr AsnTyr Asp Thr Ile Gly Lys Leu Thr Glu Phe 6110 6115 6120 Val Ala Pro LeuVal Gly Pro Ala Gly Ala Ala Pro Ala Ala Glu 6125 6130 6135 Pro Val MetAla Glu Ala Pro Gln Gln Ser Ser Ala Ser Leu Glu 6140 6145 6150 Asp LeuVal Gln Asp Val Ile Glu Arg Glu Leu Gly Arg Thr Ala 6155 6160 6165 AspPro Ala Lys Ser Phe Val Asp Asn Gly Phe Gly Ser Phe Asp 6170 6175 6180Met Leu Arg Val Val Ala Ser Leu Glu Arg Val Phe Gly Ala Leu 6185 61906195 Arg Lys Thr Leu Leu Phe Asp His Pro Thr Ile Gly Ala Leu Ala 62006205 6210 Ala His Leu Ala Glu Thr His Gly Pro Glu Ala Ala Ser Arg Leu6215 6220 6225 Ser Ser Pro Pro Gln Asp Arg Pro Arg Pro Gly Pro Ala AlaSer 6230 6235 6240 Gln Glu Pro Tyr Thr Gly Gly Ala Leu Val Val Glu LysLys Ala 6245 6250 6255 Leu Ala Gly Gln Pro Glu Leu Ala Ser Ala Val AlaGly Leu Glu 6260 6265 6270 Ser Ala Tyr Gly Arg Glu Gly Gly Leu Pro GlyArg Asp Ile Ala 6275 6280 6285 Pro Leu Ile Phe Leu Gly Ala Gly Arg ThrAla Tyr Phe Asn Phe 6290 6295 6300 Ser Val Arg Asp Asp Ala Met Leu ThrTrp Ser Tyr Val Gly Pro 6305 6310 6315 Thr Glu Glu Met Pro Ala Leu AlaThr Glu Phe Val Arg Tyr Gly 6320 6325 6330 Gln Ala His Gly Leu Ala AlaAsn Ile Val Ser Leu Ile Arg Leu 6335 6340 6345 Glu Glu Val Asp Gly ValArg Phe Thr Ala Thr Pro Phe Gly Ala 6350 6355 6360 Leu Gln Arg Leu GluGly Ile Lys Asp Phe Ser Leu Glu Gly Gly 6365 6370 6375 Arg Met Gln ArgLeu Arg Tyr Ala Val Arg Lys Phe Glu Lys Ala 6380 6385 6390 Gly Thr CysArg Thr Glu Glu Tyr Ala Val Gly Ser Asp Pro Arg 6395 6400 6405 Thr AspGln Glu Ile Thr Thr Leu Ile Asp Arg Trp Ser Ala Ala 6410 6415 6420 LysGlu Met Val Asn Pro Tyr Val Ser Thr Val Arg Asp Glu Ile 6425 6430 6435Gly Arg Gly Ile Leu Ala Ala Arg His Arg Met Phe Leu Thr Tyr 6440 64456450 Leu Asp Asp Arg Met Val Ser Ala Val Ile Val Thr Lys Ile Pro 64556460 6465 Ser Glu Asp Gly Tyr Leu Leu Asp Leu Glu Phe Tyr Pro Glu Asp6470 6475 6480 Ala Pro Leu Gly Ser Leu Asp His Thr Val Val Lys Ile IleGlu 6485 6490 6495 Arg Thr Ala Ala Glu Gly Cys Thr Val Phe Ser Phe GlyGly Ser 6500 6505 6510 Phe Gly Ala Lys Val Cys Glu Ser Pro Asn Ala AlaPro Glu Ala 6515 6520 6525 Glu Ala Ala Leu Thr Glu Leu Arg Ser Arg GlyIle Phe Thr Gly 6530 6535 6540 Asp Gly Asn Leu Arg Phe Lys Asn Lys PheArg Thr Glu Asn Leu 6545 6550 6555 Pro Leu Tyr Leu Cys Gln Pro Ala AspAla Glu Arg Thr Asp Val 6560 6565 6570 Ser Arg Leu Ile Leu Met Ile AlaAsn Pro Glu Val Gly Gly Asp 6575 6580 6585 Arg Ala Pro Thr Ala Pro ThrAla Leu Ala Ala Pro Ala Pro Arg 6590 6595 6600 Glu Ala Pro Ala Ala ProAla Pro Arg Gln Ala Pro Ala Pro Gly 6605 6610 6615 Gln Ala Pro Ala AlaArg Pro Lys Pro Ala Ala Ala Pro Pro Ala 6620 6625 6630 Ala Val Ala AlaGln Ala Thr Glu Val Pro Ala Asp Ala Arg Arg 6635 6640 6645 Arg Glu ArgGln Leu Ala Asp His Gly Trp Asn Ala Leu His Leu 6650 6655 6660 Ala SerGly Asp Val Glu Phe Asp Leu Ile Thr Asp Ser Trp Ala 6665 6670 6675 GluLeu Asp Arg Pro Phe Val His Ala Arg Thr Ala Arg Leu His 6680 6685 6690Ala Gly Ala Ala Gly Arg Pro Val Gly Gln Thr Leu Glu Gly Leu 6695 67006705 Asp Leu Leu Pro Phe Ser Cys Val Val Ala Thr Thr Ser Gly Arg 67106715 6720 Ala Ala Glu Ala Ala Leu Cys Arg Ala Trp Pro Gln Gln Gly Val6725 6730 6735 Val Val His Asn Ser Leu Phe Pro Thr Trp Tyr Phe Asn HisLeu 6740 6745 6750 Asp His Gly Phe Thr Pro Val Ala Ala Arg Arg Ala AlaGly Ala 6755 6760 6765 Asp Asp Gly Val Phe Arg Gly Asp Leu Asp Leu GlyHis Leu Asn 6770 6775 6780 Gly Leu Leu Thr Glu His Ala Gly Arg Ile AlaPhe Leu Cys Val 6785 6790 6795 Glu Val Ser Asn Asn Ala Gln Gly Gly AlaAla Leu Ser Leu His 6800 6805 6810 Asn Leu Thr Gly Ile Arg Glu Thr AlaAsp Arg His Gly Leu Gln 6815 6820 6825 Leu Val Leu Asp Ala Thr Arg ValLeu Asp Asn Ala Ala Leu Ile 6830 6835 6840 Ala Ala His Glu Pro Gly GlnThr Gly Arg Asp Pro Leu Asp Val 6845 6850 6855 Ala Arg Glu Leu Leu SerLeu Ala Asp Ser Val Thr Ile Ser Leu 6860 6865 6870 Ser Lys Asp Phe GlyVal Asp Thr Gly Gly Ile Val Ala Thr Asp 6875 6880 6885 Asp Pro Thr ValAla His His Leu Arg Glu Arg Ile Ala Leu Arg 6890 6895 6900 Gly Pro GluAla Gly Arg Ala Thr Arg Ala Leu Ala Ala Ala Ala 6905 6910 6915 Leu AspAsp Leu Gly Trp Ala Ala Thr Ala Thr Gly Glu Arg Val 6920 6925 6930 ArgArg Val Ala Asp Leu Arg Gln Ala Leu Ala Ala Ala Gly Ala 6935 6940 6945Pro Val Ala Pro Gly Thr Gly Thr His Cys Val Leu Leu Asp Thr 6950 69556960 Ala Arg Leu Pro Ala Leu Arg Gly His Glu His Pro Val Pro Ala 69656970 6975 Phe Leu Ala Trp Leu Tyr Leu His Thr Gly Ile Arg Ala Ala Ala6980 6985 6990 His Leu Asp Asp Gly Pro Gly Thr Ser Ser Leu Val Arg LeuAla 6995 7000 7005 Leu Pro Val Gly Leu Gly Gln Arg Glu Thr Ala Glu LeuThr Ala 7010 7015 7020 Arg Leu Thr Ala Leu Phe Gly Ala Pro Gln Gln IlePro Glu Leu 7025 7030 7035 Leu Leu Ala Ala Ser Asp Gly Pro Ala Ala LeuAla Ser Tyr His 7040 7045 7050 Pro Val Glu Gln Val Pro Asp Asp Ile ArgGlu Ala Met Ala Glu 7055 7060 7065 Gly His Thr Ala Glu Asn Asp Asn TrpAla Val Leu Arg Glu His 7070 7075 7080 His Pro Gly Val Glu Arg Val LeuLeu Arg Leu Pro Ala Gly Asp 7085 7090 7095 Gly Gly Gly Asp Val Glu ValPhe Thr Ala Gly Ala Gly Pro Ala 7100 7105 7110 Leu Leu Met Met Leu ProPhe Asn Ile Gly Ala Gly Leu Phe Gly 7115 7120 7125 Pro Gln Phe Ala AlaLeu Ser Glu Arg Tyr Arg Val Ile Val Val 7130 7135 7140 His His Pro GlyVal Gly Asp Thr Thr Ala Cys Glu Glu Leu Gly 7145 7150 7155 Tyr Glu GlyIle Ala Asp Leu Cys Leu Arg Ala Leu Arg Arg Leu 7160 7165 7170 Gly ValGln Gly Pro Val His Val Ala Gly Ala Ser Phe Gly Gly 7175 7180 7185 IleThr Ala Gln Thr Phe Ala Leu Arg His Pro Glu Ser Thr Ala 7190 7195 7200Ser Leu Thr Leu Ile Gly Ser Ser Tyr Lys Leu Gly Asn Arg Ala 7205 72107215 Gly Glu Val Asn Arg Leu Ala Leu Val Ala Lys Glu Asp Phe Asp 72207225 7230 Gln Val Gln Ser Leu Ser Gly Ser Ser Arg Leu Asp Arg Glu Arg7235 7240 7245 Ala Arg Phe Glu Arg Leu Leu Leu Arg Cys Glu Ser Met AspPro 7250 7255 7260 Gln Thr Gly Leu Arg Tyr Leu Asp Val Phe Ala Thr AlaPro Asp 7265 7270 7275 Leu Leu Gly Arg Leu Gly Asp Ile Ala Val Pro ThrLeu Ile Val 7280 7285 7290 Gln Gly Arg His Asp Thr Val Ile Pro Gln LysThr Ala His Leu 7295 7300 7305 Leu His Gly Ala Ile Pro Asp Ala Arg TyrHis Glu Val Pro Asp 7310 7315 7320 Ala Gly His Phe Pro Ser Leu Ser SerSer Glu Glu Phe Asn Ala 7325 7330 7335 Val Leu Ser Ala Phe Leu Glu GluHis Pro Ala 7340 7345 47 319 PRT Streptomyces atroolivaceus 47 Met ThrIle Thr Ser Ser Leu Asp Val Arg Pro Glu Ile Lys Gln Ala 1 5 10 15 ValThr Val Arg Pro Gly Met Cys Gly Pro Gly Ser Leu Phe Val Gly 20 25 30 GlnLeu Gly Asp Trp Thr Trp Glu Thr Val Ser Ala Gln Cys Asp Thr 35 40 45 AspVal Phe Ala Ala Arg Asp Ala Ser Gly Asn Pro Thr Tyr Leu Ala 50 55 60 PheTyr Tyr Phe Arg Val Arg Gly Gly Arg Glu Leu His Pro Gly Ser 65 70 75 80Leu Thr Phe Gly Asp Arg Leu Thr Val Thr Ser Gly Cys Tyr Asp Gln 85 90 95Gly Thr Glu Ser Val Leu Thr Leu His Arg Ile Asp Arg Ala Gly Ser 100 105110 Asp Asp Ala Gln Arg Pro Leu Asp Leu His Glu Phe Tyr Glu Arg Pro 115120 125 Arg Asp Gly Ser Leu Tyr Val Glu Asn Phe Asn Arg Trp Val Thr Arg130 135 140 Ser Ala Pro Gly Ser Asn Glu Asp Leu Val Lys Ser Ser Pro ProGly 145 150 155 160 Phe Arg Asn Asp Gly Leu Pro Gln Leu Pro Ala Ala TyrSer Pro Arg 165 170 175 Ala Val Tyr Arg Glu Ala Arg Thr Ala His Thr PheArg Ala Leu Asp 180 185 190 Glu Pro Gly Phe Arg Leu Leu Pro Asp Thr ValGlu Val Glu His Pro 195 200 205 Val Asp Ile Val Arg Asp Val Asn Gly ValGly Leu Leu Tyr Phe Ala 210 215 220 Ser Tyr Phe Ser Met Val Asp Lys AlaAla Leu Ala Leu Trp Arg Arg 225 230 235 240 Leu Gly Arg Ser Asp Arg AlaPhe Leu Arg Arg Val Val Val Asp Gln 245 250 255 Gln Met Cys Tyr Leu GlyAsn Ala Asp Leu Asp Ser Val Leu Thr Leu 260 265 270 Gly Ala Arg Val ArgVal Ser Thr Glu Thr Pro Gly Glu Glu Leu Val 275 280 285 Asp Val Val IleSer Asp Arg Asp Ser Gly Arg Val Ile Ala Val Ser 290 295 300 Thr Leu HisThr Gln His Asp Ala His Asp Pro Lys Gly Glu Ala 305 310 315 48 86 PRTStreptomyces atroolivaceus 48 Met Thr Gln Ala Pro Leu Asp Val Arg AlaGln Thr Ala Arg Leu Val 1 5 10 15 Val Glu Val Val Thr Glu Ile Leu ProGly Val Asp Pro Gln Leu Ile 20 25 30 Gly Gly Lys Arg His Leu Lys Asp LeuGly Ala Asp Ser Val Asp Arg 35 40 45 Val Glu Ile Ile Ala Ala Leu Leu AspArg Thr Arg Val Asp Ala Pro 50 55 60 Met Ser Asp Phe Ser Asp Leu Pro AspIle Asp Ser Leu Ile Asp Phe 65 70 75 80 Leu Ser Gly Ala Pro Arg 85 49416 PRT Streptomyces atroolivaceus 49 Met Thr Ala Thr Gly Ala Glu GlnAla Tyr Gly Ile Glu Ala Leu Asn 1 5 10 15 Val Trp Cys Gly Leu Ala ArgLeu Thr Ala Ala Asp Leu Phe Ala Gly 20 25 30 Arg Gly Leu Asp Pro Glu ArgLeu Asp Asn Leu Met Met Ser Glu Arg 35 40 45 Ser Ile Gly Leu Pro Ile GluAsp Pro Val Thr Asn Ala Val Asn Ala 50 55 60 Ala Arg Pro Leu Ile Glu AlaLeu Ser Pro Glu Glu Arg Ala Arg Ile 65 70 75 80 Glu Leu Val Val Thr SerThr Glu Ser Gly Val Asp Tyr Ser Lys Ser 85 90 95 Leu Thr Ser Tyr Val HisLys Tyr Leu Gly Leu Asn Arg His Cys Arg 100 105 110 Leu Ile Glu Val LysGln Ala Cys Phe Gly Ala Thr Ala Ala Val Gln 115 120 125 Thr Ala Val GlyTyr Leu Ala Ser Gly Ile Ser Pro Gly Ala Lys Ala 130 135 140 Leu Val IleAla Thr Asp Val Ala Val Val Asp Glu Lys Ala Glu Tyr 145 150 155 160 SerGlu Pro Ala Ala Gly His Gly Ala Ala Ala Met Leu Leu Ser Asp 165 170 175Arg Pro Arg Val Leu Ala Met Asp Leu Gly Ala Phe Gly Asn Tyr Ser 180 185190 Tyr Glu Thr Leu Asp Ser Ala Arg Pro Ser Pro Arg Phe Asp Ile Ala 195200 205 Asp Val Asp Arg Ser Leu Phe Ala Tyr Leu Asp Cys Leu Lys Asn Ala210 215 220 Tyr Ala Asp Tyr Ala Ala Arg Val Thr Asp Val Asp Phe Thr ArgAsp 225 230 235 240 Phe Asp His Leu Val Met His Thr Pro Phe Ala Gly LeuVal Lys Ala 245 250 255 Gly His Arg Lys Met Met Arg Glu Gln Gly Val ThrGly Pro Arg Ile 260 265 270 Asp Glu Asp Phe Ala Arg Arg Val Ala Pro SerLeu Ile Tyr Pro Gly 275 280 285 Ser Val Gly Asn Leu Cys Ser Gly Ser ValTyr Leu Ala Leu Ala Ser 290 295 300 Leu Leu Asp Ser Gly Val Val Thr AlaPro Ser Arg Val Gly Leu Phe 305 310 315 320 Ser Tyr Gly Ser Gly Cys SerSer Glu Phe Phe Ser Gly Ile Val Asp 325 330 335 Glu Gln Ser Ala Ala ThrVal Ala Glu Gln Gly Ile Gly Lys Arg Leu 340 345 350 Glu Ala Arg Ala ArgIle Thr Phe Asp Glu Tyr Leu Ala Val Leu Glu 355 360 365 His Asn Leu GluCys Leu Val Pro Val Glu Asn Arg Thr Val Asp Pro 370 375 380 Ala Glu TrpGlu Pro Leu Leu Asp Arg Val Gly Asp Arg Pro Glu Ile 385 390 395 400 LeuThr Phe Thr Gly Val Lys Asp Tyr His Arg Gln Tyr Ala Trp Arg 405 410 41550 267 PRT Streptomyces atroolivaceus 50 Met Tyr Gly Ser Arg Thr Ala ThrArg Pro Ala Gly Glu Asp Gly Ala 1 5 10 15 Ala Ala Gly Leu Arg Leu PheCys Phe Ala His Ala Gly Gly Gly Ser 20 25 30 Ser Phe Phe His Pro Trp ArgArg Ala Leu Gly Pro Gly Val Asp Val 35 40 45 Arg Pro Val Val Leu Pro GlyArg Glu Arg Arg Ala Arg Glu Thr Ser 50 55 60 His Thr Arg Met Gly Pro LeuVal Glu Gly Leu Val Thr Glu Leu Ala 65 70 75 80 Pro Gln Leu Asp Leu ProTyr Val Leu Phe Gly His Ser Leu Gly Ser 85 90 95 Ile Val Ala Tyr Glu ThrAla Arg Ala Leu Leu Glu Arg Gly Ser Arg 100 105 110 Pro Pro Leu Ala LeuLeu Val Ser Gly Arg Arg Gly Pro Phe Val Pro 115 120 125 Asp His Arg ArgPro Val His Asn Leu Pro Glu Asp Glu Phe Leu Ala 130 135 140 Glu Val SerArg Leu Gly Gly Thr Pro Ser Glu Val Leu Arg Gln Arg 145 150 155 160 AspLeu Leu Arg His Phe Leu Pro Pro Leu Arg Ala Asp His Glu Val 165 170 175Asn Glu Thr Tyr Arg Pro Val Pro Pro Pro Gly Pro Ala Leu Thr Cys 180 185190 Pro Val Phe Ala Phe Thr Gly Asp Ala Asp Pro Leu Ala Asp Pro His 195200 205 Ala Val Ala Arg Trp Arg Glu Val Thr Ser Gly Asp Phe Arg Leu Arg210 215 220 Val Phe Pro Gly Asp His Phe Tyr Leu Lys Gly Ala Pro Asp AspLeu 225 230 235 240 Met Ser Ala Leu Arg Ala Ala Met Gly Ser Val His GlnGly Ser Ser 245 250 255 Val Leu Arg Arg Gln Val His Lys Val Ala Thr 260265 51 227 PRT Streptomyces atroolivaceus 51 Met Asn Leu Leu Asp Val ThrGlu Pro Thr Gly Phe Asp Ala Arg Leu 1 5 10 15 Arg Asp Phe Leu Arg AspGln Ser Pro His Thr Thr Thr Thr Met Val 20 25 30 Arg Arg Asn Gln Ser AlaTyr Ser Cys Gly Gly Gln Asp Arg Asn Val 35 40 45 Tyr Phe Leu Glu Ser GlyArg Leu Lys Thr Val Met Phe Ser Arg Ser 50 55 60 Gly Lys Glu Cys Leu LeuArg Ile His Thr Pro Gly Ser Ile Phe Gly 65 70 75 80 Glu Leu Cys Ser LeuGly Gly Ile Arg Glu Glu Ser Ala Ile Ala Met 85 90 95 Arg Asp Ser Val ValHis Arg Met Ser Tyr Glu His Phe Leu Ser Ser 100 105 110 Ile Thr Glu AlaGly Ile Val Asp Glu Phe Ile Asn His Leu Thr Arg 115 120 125 Arg Leu AlaGlu Gln Gln Gln Ser Ile Thr His Leu Val Thr Val Asp 130 135 140 Ser GluGln Arg Leu Gly Glu Ile Leu Leu Asp Leu Ala Arg Lys Leu 145 150 155 160Gly Arg Gly Asp Gly Arg Arg Pro His Ile Ala Glu Arg Ile Thr His 165 170175 Glu Glu Leu Ala Gly Met Val Gly Thr Thr Arg Ser Arg Val Gly His 180185 190 Phe Leu Lys Gly Phe Arg Asp Arg Gly Leu Val Glu Val Thr Arg Glu195 200 205 Ser His Leu Ile Ile Asp Gln Ser Arg Leu Ala Ala Tyr Leu GluSer 210 215 220 Arg Cys Val 225 52 82 PRT Streptomyces atroolivaceus 52Met Trp Asp His Lys Phe Glu Thr Leu Leu Arg Asp Arg Leu Pro Gly 1 5 1015 Leu Asp Ala Asp Ala Pro Leu Thr Glu Asp Leu Pro Leu Ala Ala Phe 20 2530 Gly Leu Asp Ser Met Ala Thr Val Gly Leu Ile Val Ala Ile Glu Glu 35 4045 Asp Tyr Gly Thr Pro Leu Pro Asp Asp Ala Val Thr Pro Ala Asn Phe 50 5560 Ile Thr Pro Gly Thr Val Trp Glu Leu Val Ser Ser Leu Asp Gly Ala 65 7075 80 Gly Arg 53 516 PRT Streptomyces atroolivaceus 53 Met Ser Gly AlaLys Leu Leu His His Trp Phe Leu Asp Gly Leu Ala 1 5 10 15 Arg Asn ProArg Gly Thr Ala Leu Arg Ile Asn Asp Arg Ser Trp Thr 20 25 30 Tyr Ala GluAla Asp Arg Thr Ala Arg Ser Trp Ala Ala Ala Leu His 35 40 45 Ser Gly GlyArg Ala Pro Arg Arg Val Gly Val Leu Ala Ala Lys Ser 50 55 60 Glu Glu SerLeu Leu Gly Phe Leu Ala Ala Leu Tyr Ala Gly Cys Thr 65 70 75 80 Ala ValPro Leu Asn Pro Glu Tyr Pro Val Gly Arg Asn Arg Asp Ile 85 90 95 Ala AlaAla Ala Gly Leu Asp Ala Leu Ile Val Asp Pro Ser Gly Ala 100 105 110 AlaGln Leu Asp Glu Val Ala Ala Ala Ala Pro Pro Leu Val Thr Leu 115 120 125Ala Pro Arg Leu Gly Glu Trp Ser Gly Pro Arg Pro Gly Thr Pro Val 130 135140 Leu Met Pro Asp Pro Ala Gly Asp Arg Pro Pro Ala Gln Gly Pro Asp 145150 155 160 Ser Leu Ala Tyr Ile Leu Phe Thr Ser Gly Ser Thr Gly Arg ProLys 165 170 175 Gly Val Pro Ile Arg His Gly Asn Val Ser Ala Phe Leu AlaAla Ser 180 185 190 Leu Pro Arg Tyr Asp Phe Gly Pro Asp Asp Val Phe GlyGln Val Tyr 195 200 205 Glu Leu Thr Phe Asp Leu Ser Met Phe Glu Val TrpCys Ala Trp Ser 210 215 220 Ser Gly Ala Cys Leu Thr Val Leu Asn Arg LeuGln Ala Leu Asn Pro 225 230 235 240 Gly Arg Tyr Ile Arg Ala His Gly IleThr Val Trp Thr Ser Thr Pro 245 250 255 Ser Leu Val Ala Ala Leu Arg ThrArg Gly Leu Leu Gly Gly Asn Ser 260 265 270 Leu Pro Ser Val Arg His ThrVal Phe Cys Gly Glu Pro Leu Pro Glu 275 280 285 Glu Ser Ala Ala Tyr TrpSer Ala Ala Ala Pro Gly Thr Ser Ile Asp 290 295 300 Asn Leu Tyr Gly ProThr Glu Leu Thr Ile Ala Cys Thr Ala Tyr His 305 310 315 320 Trp Val ProSer Pro Gly Ser Gly Thr Val Pro Ile Gly Glu Pro Asn 325 330 335 Thr GlyLeu Arg Tyr Val Leu Leu Asp Asp Gly Gln Val Gly Ala Asp 340 345 350 ThrGly Glu Leu Cys Val Thr Gly Pro Gln Met Phe Asp Gly Tyr Leu 355 360 365Asp Pro Ala Asn Asp Thr Asp Arg Phe Leu Thr His Asp Gly Gln Arg 370 375380 Trp Tyr Arg Thr Gly Asp Arg Val Arg Arg Thr Pro Asp Gly Thr Leu 385390 395 400 Val His Leu Gly Arg Asp Asp Gly Gln Val Lys Ile His Gly TyrArg 405 410 415 Val Glu Leu Ser Glu Val Glu Glu Ala Val Arg Ala Cys AlaPro Gly 420 425 430 Thr Glu Ala Val Val Leu Ala Val Pro Gly Ala Ala GlyPro Val Leu 435 440 445 Ala Ala Phe Val Ile Gly Asp Asp Ala Ala Gly ArgLeu Thr Gly Thr 450 455 460 Ala Gln Asp Leu Ala His Arg Leu Pro Pro TyrMet Leu Pro Ala His 465 470 475 480 Leu Trp Ala Leu Ala Asp Pro Pro LeuAsn Pro Asn Gly Lys Thr Asp 485 490 495 Arg Leu Arg Leu Arg Glu Glu AlaAla Arg Arg Leu Gly Arg Asn Leu 500 505 510 Ala Pro Ala Ser 515 54 575PRT Streptomyces atroolivaceus 54 Met Ser Ala Val Phe Asp Ala Arg SerAsp Leu Ala Val Pro Ala Pro 1 5 10 15 Ala Arg Pro His Gly Gln Gly ArgAla Asp Ala Ala Pro Glu Val Ile 20 25 30 Ser Val Thr Asp Leu Thr Val AlaAsp Pro Asn Gly Arg Pro Val Leu 35 40 45 Asp Gly Leu Asn Leu Thr Val ArgAla Gly Glu Arg Val Gly Val Val 50 55 60 Gly Glu Ser Gly Ser Gly Lys ThrThr Leu Ala Leu Ala Leu Leu Gly 65 70 75 80 Ala Leu Arg Pro Gly Leu ArgThr Ala Gly Gly Gly Val Arg Val Ala 85 90 95 Gly Glu Asp Pro Leu Ala LeuArg Gly Arg Asp Leu Arg Arg Leu Arg 100 105 110 Arg His Thr Val Ala TyrLeu Pro Gln Asp Pro Ala Ala Ala Leu Thr 115 120 125 Pro Thr Met Arg ValGly Ala Leu Val Ser Glu Leu Ala Val Arg Arg 130 135 140 Thr Pro Glu AspAla Ala Arg Thr Leu Arg Ser Val Gly Leu Pro Asp 145 150 155 160 Gly ArgAsp Phe Leu Arg Arg Phe Pro His Glu Leu Ser Gly Gly Gln 165 170 175 GlnGln Arg Leu Ala Leu Gly Arg Val Leu Ala Ala Gly Pro Ala Ala 180 185 190Leu Val Leu Asp Glu Pro Thr Thr Gly Leu Asp Val Leu Thr Gln Arg 195 200205 Leu Val Leu Asp Glu Val Ala Arg Leu Ala Ala Glu Arg Asn Leu Thr 210215 220 Leu Leu Phe Ile Thr His Asp Leu Pro Ala Ala Ala Arg Leu Thr Asp225 230 235 240 Arg Leu Ala Val Leu Lys Ala Gly Arg Val Val Glu Glu GlyPro Val 245 250 255 Asp Ala Val Leu Gly Arg Pro Arg Ala Ala Tyr Thr ArgGlu Leu Val 260 265 270 His Ala Val Pro Asp Ile Gly Glu Ala Ala Ala ArgPro Ala Ala Pro 275 280 285 Ala Gly Gly Ala Ala Asp Thr Ala Pro Val LeuArg Val Ser Gly Leu 290 295 300 Arg Ala Gly His Gly Arg Gly Arg Ala ArgVal Thr Thr Ala Gln Asp 305 310 315 320 Val Ser Phe Arg Ile Glu Pro GlyGlu Cys Val Ala Leu Leu Gly Gly 325 330 335 Ser Gly Thr Gly Lys Thr ThrLeu Ala Arg Cys Val Thr Gly His His 340 345 350 Ser Pro Asp Gly Gly LeuIle Glu Leu Ala Gly Ser Ala Val Pro Gly 355 360 365 Pro Leu Arg Gly ArgThr Leu Asp Gln Arg Arg Leu Val Gln Leu Val 370 375 380 Ala Gln Asp AlaAsn Gly Ser Leu Asn Pro Arg Arg Thr Val Gly Ala 385 390 395 400 Ala IleAla Arg Pro Leu Arg Val Leu Arg Gly Leu Gly Gly Glu Ala 405 410 415 AlaAla Ala Glu Thr Glu Arg Leu Leu Arg Leu Val Gly Leu Asp Pro 420 425 430Gln Trp Ala Gly Arg Leu Pro Arg Thr Leu Ser Gly Gly Gln Arg Gln 435 440445 Arg Val Ala Ile Ala Arg Ala Leu Ala Ala Gln Pro Arg Leu Leu Leu 450455 460 Cys Asp Glu Val Thr Ser Ser Leu Asp Val Arg Ile Gln Ala Glu Ile465 470 475 480 Leu Ala Leu Leu Asp Gly Leu Arg Ser Arg Leu Gly Leu GlyLeu Leu 485 490 495 Leu Ile Ser His Asp Leu Gly Val Ile Ala Arg Met AlaAsp Arg Val 500 505 510 Leu Val Leu Asp Gly Gly Thr Val Arg Glu Gln GlyPro Val Ala Glu 515 520 525 Val Phe Ala Ala Pro Arg His Pro Trp Thr ArgAla Val Leu Asp Ala 530 535 540 Val Pro Ser Val Arg Gly Glu Leu Ala GlyArg Ala Glu Arg Thr Pro 545 550 555 560 Ala Ala Asp Glu Ser Arg Pro ArgGlu Thr Ala Val Asp Gly Val 565 570 575 55 287 PRT Streptomycesatroolivaceus 55 Met Thr Ala Pro Thr Pro Thr Ala Thr Glu Ala Leu Thr ProAla Gly 1 5 10 15 Thr Gln Ala Pro Asp Thr Gly Pro Ala Ala Pro Arg ProGly Arg Arg 20 25 30 Ala Ala Leu Leu Leu Leu Leu Pro Val Ala Ala Val ValLeu Phe Ala 35 40 45 Leu Ile Gly Pro Trp Leu Ala Pro His Asp Pro Ala ArgIle Thr Ala 50 55 60 Ala Pro Tyr Asp Thr Gly His Leu Leu Leu Gly Ser AspGly Asp Gly 65 70 75 80 Arg Asp Val Trp Ser Arg Phe Leu Ala Gly Gly ArgPro Leu Val Leu 85 90 95 Ile Pro Leu Leu Ala Thr Ala Val Thr Thr Leu LeuGly Thr Ala Ser 100 105 110 Gly Val Leu Ala Gly Tyr Ala Gly Gly Arg ValAsp Ala Val Val Ser 115 120 125 Arg Leu Asp Thr Leu Leu Leu Ala Leu ProPro Val Leu Val Leu Leu 130 135 140 Val Leu Met His Gly Trp Gly Tyr ArgThr Leu Thr Leu Val Thr Ala 145 150 155 160 Val Ala Val Thr Gly Val ProPhe Val Ser Arg Val Ala Arg Ala Ala 165 170 175 Thr Ala Gln Val Val HisAla Gly Tyr Val Asp Gln Ala Val Ala Leu 180 185 190 Gly Asp Gly Pro ValAla Val Met Ala Arg Glu Ile Val Pro Asn Ile 195 200 205 Val Arg Pro ValLeu Ala Asp Ala Gly Thr Arg Leu Ala Ile Ala Ile 210 215 220 Thr Leu ThrAla Ser Ala Gly Phe Leu Gly Phe Gly Pro Asp Ser Pro 225 230 235 240 AsnTrp Gly Ala Met Ile Ser Glu Asn Met Glu Gly Val Thr Leu Ala 245 250 255Pro Trp Gly Val Ala Ala Pro Ala Leu Gly Leu Ala Val Leu Thr Val 260 265270 Ser Val Asn Leu Ala Leu Asp Arg Ala Ile Ala Arg Phe Val Pro 275 280285 56 321 PRT Streptomyces atroolivaceus 56 Met Thr Ser Ala Val Arg LeuAla Ala Leu Ala Val Arg Arg Leu Gly 1 5 10 15 Gly Ala Leu Ala Thr LeuLeu Val Gly Ser Phe Val Leu Phe Ala Ala 20 25 30 Ala Glu Val Leu Pro GlyAsp Ala Ala Ala Ala Ala Leu Gly Gly Ser 35 40 45 Ala Thr Pro Ala Asp ValAla Arg Met His Ala Glu Leu Gly Leu Asp 50 55 60 Arg Pro Val Leu Val ArgTyr Gly Asp Trp Ile Gly Ala Ala Phe Ser 65 70 75 80 Gly Asp Leu Gly HisSer Leu Val Asp Arg Arg Pro Val Ala Glu Ile 85 90 95 Ile Ser Gly Pro LeuAla Asp Thr Leu Leu Leu Ala Gly Val Cys Ala 100 105 110 Leu Leu Thr ValVal Leu Ala Leu Ala Val Gly Ile Ala Ala Gly Leu 115 120 125 Arg Pro GlyGly Leu Leu Asp Arg Leu Leu Ser Gly Gly Ala Val Thr 130 135 140 Leu ValSer Val Pro Gln Phe Val Thr Ala Gly Ile Leu Thr Leu Val 145 150 155 160Phe Ser Ser Trp Leu Gly Val Leu Pro Ala Val Ser Leu Val Pro Phe 165 170175 Gly Glu Pro Leu Leu Ala His Pro Glu Ala Leu Val Leu Pro Val Ala 180185 190 Ala Leu Thr Leu Phe Ser Ala Ser Trp Ala Ser Arg Met Val Arg Ala195 200 205 Ala Val Val Asp Ala Ala Ala Ala Pro His Val Glu Ala Ala ArgLeu 210 215 220 Ala Gly Leu Ser Pro Ala Arg Val Val Leu Arg His Leu LeuPro Thr 225 230 235 240 Ala Ala Gly Pro Cys Ala Gln Val Phe Gly Trp LeuAla Gly Thr Leu 245 250 255 Val Gly Gly Thr Ala Val Val Glu Lys Val PheAsn Tyr Pro Gly Leu 260 265 270 Ser Arg Glu Leu Val Ala Ala Val Gln HisHis Asp Ala Ala Val Leu 275 280 285 Glu Gly Ile Gly Leu Leu Met Ala AlaAla Val Val Gly Gly Leu Leu 290 295 300 Leu Ala Asp Leu Leu Ala Met AlaVal Asp Pro Arg Met Arg Thr Ala 305 310 315 320 Thr 57 513 PRTStreptomyces atroolivaceus 57 Met Ser Arg Val Asn Gly Val Gly Arg ArgArg Phe Leu Gln Trp Gly 1 5 10 15 Ala Ala Ala Ala Thr Ala Thr Ala GlyAla Thr Ala Leu Ser Gly Cys 20 25 30 Ala Thr Ser Lys Ser Gly Asp Thr ArgSer Val Gly Ser Ala Gly Lys 35 40 45 Leu Leu Arg Val Gly Ala Ser Asp AlaThr Gly Thr Thr Ser Leu Asp 50 55 60 Pro Arg His Ala Gly Phe Gly Ala SerTyr Ile Ala Leu Tyr His Leu 65 70 75 80 Tyr Asp Gln Leu Met Tyr Leu LysGly Ser Arg Tyr Glu Leu Gly Leu 85 90 95 Ala Glu Ser Val Glu Pro Asn LysAsp Ala Ser Glu Trp Ile Val Arg 100 105 110 Leu His Lys His Ala Val PheHis Asp Gly Ser Pro Val Arg Ala Ala 115 120 125 Asp Val Ala Tyr Ser LeuArg Thr Leu Ala Thr Pro Pro Ser Thr Arg 130 135 140 Ala Asn Asn Phe ValAsp Leu Asp Leu Asp Arg Leu Lys Ala Val Asp 145 150 155 160 Ala His ThrLeu Lys Val Gly Leu Lys Arg Pro Arg Gly Asp Phe Arg 165 170 175 Glu AlaVal Leu Ala Val Phe Ser Pro Val Phe Pro Glu Gly Thr Arg 180 185 190 GlyLys Asp Phe Asp Lys Gly Val Gly Ser Gly Pro Tyr Arg Leu Glu 195 200 205Glu His Asp Gly Lys Asn Leu Arg Leu Ala Val Asn Asp Lys Tyr Trp 210 215220 Gly Gly Thr Ser Tyr Phe Arg Arg Ile Glu Leu Asp Leu Ile Ala Asp 225230 235 240 Pro Ala Ala Arg Leu Ala Ala Leu Lys Ser Gly Gln Ile Asp TyrAla 245 250 255 Phe Gly Ile Thr Ala Thr Gly Ala Lys Ala Glu Arg Gly AsnSer Ala 260 265 270 Ile Arg Val Arg Arg Gly Gly Thr Ala Asn Ser Asn AlaLeu Ser Phe 275 280 285 Ala Met Asn Glu Gln Leu Ala Pro Phe Asp Asn ProAsp Val Arg Arg 290 295 300 Ala Val Arg Leu Ala Ala Asp Arg Glu Ala LeuVal Arg Gln Val Leu 305 310 315 320 Leu Gly Leu Gly Thr Lys Gly Asp AspVal Val Gly Lys Gly Leu Pro 325 330 335 Gly Tyr Ala Gly Gly Asp Arg LysArg Asp Val Ala Glu Ala Arg Arg 340 345 350 Leu Phe Ala Arg Ala Lys ValHis Ser Leu Thr Leu His Ala Ala Asp 355 360 365 Thr Val Pro Gly Phe ValAla Ala Ala Arg Leu Leu Gly Gln Gln Leu 370 375 380 Lys Glu Ala Gly ValGlu Leu Asp Val Gln Lys Val Pro Ala Asp Thr 385 390 395 400 Tyr Tyr AlaAsp Leu Lys Gly Leu Ala Arg Thr Pro Phe Gln Thr Phe 405 410 415 Tyr TyrAla Asn Arg Pro Ala Ala Val His Leu Ser Ala Thr Thr Thr 420 425 430 LysGly Ala Phe Phe Asn Val Thr Gly Leu Gly Glu Pro His Phe Lys 435 440 445Arg Leu Ala Ser Ala Gln Ala Thr Val Asp Asp Ser Lys Arg Glu Ala 450 455460 Ala Phe Ala Ala Val Gln Arg Gly Leu Tyr Ser Ser Gly Gly Asp Leu 465470 475 480 Leu Trp Gly Phe Gln Glu Gln Leu Asp Ala Ser Ala Pro Gly LeuSer 485 490 495 Gly Val Trp Ser Ser Gln Ser Ile Pro Val Phe Thr Arg AlaArg Ser 500 505 510 Ala 58 120 PRT Streptomyces atroolivaceus 58 Met SerThr Asp Thr Glu Ile Pro Ile His Pro Leu Phe Ala Glu Met 1 5 10 15 IleArg Cys Thr Ala Glu Arg Asp Leu Asp Gly Leu Val Asp Leu Tyr 20 25 30 HisPro Glu Ala Glu Trp Ile Arg Phe Thr Gly Pro Val Arg Gly Arg 35 40 45 GluAsp Ile Arg Glu Leu Leu Lys Arg Tyr Trp Glu Leu Asp Leu Gln 50 55 60 HisVal Asp Met Asn Glu Phe Ile Gln Thr Asp Asp Thr Val Met Met 65 70 75 80Arg Gly Thr Met Ile Val Arg Gly Glu Thr Val Val Thr Phe Gly Val 85 90 95Tyr Val Leu Arg Glu Gly Lys Ile Trp Arg Gln Cys Gly Ala Asp Glu 100 105110 Gly Gly Thr Arg Asp Trp Trp Ala 115 120 59 516 PRT Streptomycesatroolivaceus 59 Met Thr Glu Arg Thr Phe Ser Tyr Leu Thr Asp Gln Leu ArgSer His 1 5 10 15 Ala Ala Leu His Pro Asp Arg Thr Ala Leu Val Ile AspGly Cys Pro 20 25 30 Asp Leu Leu Tyr Gly Glu Trp Asp Arg Arg Ser Glu AlaLeu Ala Arg 35 40 45 Gly Leu Leu Ala Ala Gly Thr Ser Arg Gly Thr Arg IleGly Ile Phe 50 55 60 Phe Gly Gly Met Asp Trp Ala Gly Tyr Ala Val Ala TyrLeu Gly Ala 65 70 75 80 Leu Lys Ala Gly Ala Thr Val Leu His Leu Pro LeuAla Leu Pro Ala 85 90 95 Asp Glu Leu Glu Arg Arg Ala Leu Gln Cys Glu LeuAla Gly Ile Val 100 105 110 His Gly Arg Thr Ala Pro Pro Thr Thr Ser AlaVal Ala Trp Thr Gly 115 120 125 Thr Leu Asp Glu Leu Ser Ala Pro Gly GluThr Pro Val Asp Leu Val 130 135 140 His Ser Pro Ala Asp Ala Ala Glu IleVal Tyr Ser Ser Gly Thr Thr 145 150 155 160 Gly Leu Ala Arg Gly Val ValVal Ser His Gln Asn Leu Ala Thr Ala 165 170 175 Gly Gly Pro Pro Ser ValMet Ala His Asp Glu Pro Thr Pro Met Val 180 185 190 Ala Ser Val Asn LeuGly Ile Thr Ala Ser Ala Thr Thr Val Ser Met 195 200 205 Val Leu Asn AlaThr Pro Thr Thr Leu Val Leu Ala Pro Pro Gly Asp 210 215 220 Ala Asp ArgLeu Cys Ala Leu Ile Glu His His Ala Ala Ser Thr Val 225 230 235 240 MetMet Thr Pro Asn Leu Ala Val Gln Met Thr Arg Asp Gly Ala Leu 245 250 255Gly Arg Tyr Asp Leu Thr Ser Val Thr Thr Val Ala Thr Ala Ser Ala 260 265270 Phe Leu His Pro Pro Leu Ala Arg Ala Leu Leu Ala Ala Met Pro Arg 275280 285 Ala Arg Val Ile Gly Ala Tyr Ser Ala Ser Gln Ala Lys Pro Ala Val290 295 300 Thr Ile Gly Thr Phe Asp Pro Ala Arg Pro Met Ser Ala Gly ArgPro 305 310 315 320 Ala Pro Gly Thr His Val Leu Ile Thr Asp Glu His GlyAla Glu Leu 325 330 335 Pro Ala His Arg Val Gly Arg Ile Trp Leu Arg AlaAsp Gly Ala Pro 340 345 350 Pro Arg Asn Arg Leu Asp Ala Gly Pro Glu AlaThr Gly Val Pro Glu 355 360 365 Gly Gly Trp Cys Asp Thr Gly Asp Leu GlyHis Val Asp Asp Glu Gly 370 375 380 Glu Leu Tyr Leu Phe Asp Arg Glu ThrAsp Ala Val Pro Thr Pro Ala 385 390 395 400 Gly Leu Val Ser Ser Leu ArgVal Glu Ser Val Leu Leu Glu His Glu 405 410 415 Ala Val Ala Asp Ala AlaVal Val Ala Ala Gly Pro Ala Gly Val Ala 420 425 430 Ala Ala Ile Val ProAla Ala Gly Ala Thr His Asp Pro Lys Leu Leu 435 440 445 Ala Ala Thr LeuAla Ala His Ala Lys Asp Ser Leu Ala Pro His Glu 450 455 460 Ile Pro GluArg Val Leu Val Val Asp Glu Leu Pro Arg Asn Asp Leu 465 470 475 480 GlyLys Val Val Lys Arg Leu Ile Arg Asp Arg Leu Thr Ala Ser Ala 485 490 495Ala Gly Val Thr Pro Ala Pro Thr Ala Ser Ala Glu Leu Asp Ala Thr 500 505510 Gly Ala Pro Ser 515 60 243 PRT Streptomyces atroolivaceus 60 Met AlaAsp Thr Leu Leu Glu Leu Pro Asp Asp Phe Ser Arg Val Leu 1 5 10 15 AlaIle Val Ala His Pro Asp Asp Ile Glu Phe Gly Ala Gly Pro Ala 20 25 30 ValAla Gln Trp Thr Ala Gln Gly Arg Glu Val Ala Tyr Leu Leu Val 35 40 45 ThrArg Gly Glu Ala Gly Ile Ser Asp Leu Glu Pro Ala Gln Cys Gly 50 55 60 ProVal Arg Glu Ala Glu Gln Arg Lys Ala Ala Ala Glu Leu Gly Val 65 70 75 80His Glu Val Asp Phe Leu Asp His Tyr Asn Asp Gly Thr Ile Glu Tyr 85 90 95Gly Pro Gly Leu Arg Arg Asp Leu Ala Arg Ala Val Arg Arg His Arg 100 105110 Pro Glu Leu Ile Val Thr Phe Asn His His Asp Thr Trp Ala Ser Gly 115120 125 Ala Trp Asn Thr Pro Asp His Arg Ala Val Gly Leu Ala Ala Leu Asp130 135 140 Ala Val Ala Asp Ala Ala Asn Arg Trp Ile Phe Pro Glu Leu LeuAsp 145 150 155 160 Glu Gly Leu Glu Pro Trp Arg Ala Gly Lys Val Ala IleAla Gly Ser 165 170 175 Pro His Ala Thr His Ala Val Ala Val Asp Asp AspSer Arg Asp Arg 180 185 190 Ala Val Arg Ser Leu Ala Ala His Asp Arg TyrLeu Gly Ser Leu Ser 195 200 205 Asp Asp Pro Pro Gln Glu Arg Ala Arg PheIle Leu Gly His Leu Leu 210 215 220 Ala Ala Thr Ala Pro Arg Phe Gly GlyArg Asp Gly Val Ala Phe Gln 225 230 235 240 Ile Val Gly 61 474 PRTStreptomyces atroolivaceus 61 Met Thr Val Arg Thr Asp Asp Gly Arg GlnAla Ala Glu Gln Ser Pro 1 5 10 15 Ala Thr Gly Ala Leu Val Ala Arg LeuIle Leu Val Thr Leu Leu Met 20 25 30 Ala Glu Leu Leu Ile Gln Val Asp GlnThr Ile Val Asn Val Ala Leu 35 40 45 Pro Phe Ile Gln Arg Asp Leu Asp PheThr Glu Ser Gly Leu Pro Trp 50 55 60 Val Val Asn Ala Tyr Gly Leu Leu TyrGly Gly Leu Leu Pro Leu Gly 65 70 75 80 Gly Arg Ile Gly Asp Leu Phe GlyArg Arg Arg Val Leu Ile Ile Gly 85 90 95 Val Thr Val Phe Thr Leu Ala SerVal Val Cys Gly Phe Ser Thr Asp 100 105 110 Pro Thr Val Met Val Ile AlaArg Ala Val Gln Gly Met Gly Gly Ala 115 120 125 Leu Thr Ala Pro Val ValLeu Ser Leu Ile Ile Thr Ser Phe Glu Glu 130 135 140 Gly Pro Ser Arg GlnArg Ala Phe Ala Leu Trp Gly Ser Ala Gln Ala 145 150 155 160 Ala Gly AlaLeu Phe Gly Leu Ile Val Gly Gly Leu Leu Thr Ser Gly 165 170 175 Pro GlyTrp Glu Phe Ser Phe Phe Val Ala Val Pro Ile Gly Ala Leu 180 185 190 ValVal Gly Leu Ala Ala Thr Thr Ile Lys Glu Ser Arg Ala Asp Glu 195 200 205Arg Pro Ser Ile Asp Val Ala Gly Ala Val Thr Phe Thr Ala Ala Leu 210 215220 Leu Met Thr Val Phe Ala Phe Leu Asp Thr Gln Arg Ser Trp Thr Ser 225230 235 240 Pro Leu Arg Gly Gly Leu Leu Val Val Ala Ala Val Leu Phe AlaVal 245 250 255 Phe Leu Ala Val Glu Arg Arg Gln Gly His Pro Phe Val ProLeu Gln 260 265 270 Val Phe Arg Leu Arg Asn Thr Ser Gly Ala Leu Ala IleMet Ala Ile 275 280 285 Phe Gly Ala Thr Gln Met Ser Tyr Phe Phe Phe ValThr Leu Tyr Leu 290 295 300 Gln Glu Ile Leu Gly Phe Ser Ala Leu Gln ThrGly Leu Ala Tyr Leu 305 310 315 320 Pro Leu Ile Ala Thr Leu Leu Val PheAla Gln Val Cys Met Lys Thr 325 330 335 Val Ala Arg Val Gly Leu Thr ArgMet Leu Met Thr Gly Leu Leu Cys 340 345 350 Leu Gly Leu Gly Met Gly TrpLeu Gly Leu Ala Ala Ser Ser Gly Ser 355 360 365 Phe Val Gly Thr Val LeuGly Pro Thr Ile Ile Ser Gly Ile Gly Leu 370 375 380 Gly Leu Thr Met MetPro Ala Gly Thr Leu Ala Thr Thr Gly Val Asp 385 390 395 400 Pro Ala AspAla Gly Ala Val Ser Gly Leu Phe Asn Thr Ala Ile Leu 405 410 415 Val GlyGly Ser Leu Gly Leu Ala Val Leu Thr Thr Val Thr Gln Ala 420 425 430 ValGly Gly Leu Gly Gly Thr His Gly Tyr Thr Val Ala Phe Leu Cys 435 440 445Ser Ala Gly Leu Ala Ala Ala Ala Ile Leu Ile Ala Ala Leu Val Ile 450 455460 Arg Val Pro Lys Gln Thr Asp Ala Ala Ala 465 470 62 400 PRTStreptomyces atroolivaceus 62 Met Ser Thr Glu Val Glu Thr Glu Lys ProAla Pro Val Ala Tyr Pro 1 5 10 15 Phe Thr Gly Ser Glu Gly Leu Glu LeuSer Gln Ser Tyr Ala Lys Leu 20 25 30 Phe Glu Asp Gly Asp Pro Ile Arg ValGln Leu Pro Phe Gly Glu Pro 35 40 45 Ala Trp Leu Val Thr Arg Tyr Asp AspAla Arg Phe Val Leu Thr Asp 50 55 60 Arg Arg Phe Ser Arg His Leu Ala ThrGln Arg Asp Glu Pro Arg Met 65 70 75 80 Thr Pro Arg Ala Val Pro Glu SerIle Leu Thr Met Asp Pro Pro Asp 85 90 95 His Thr Arg Leu Arg Thr Leu ValSer Lys Ala Phe Thr Pro Arg Arg 100 105 110 Ile Glu Ser Lys Arg Ala TrpIle Gly Glu Leu Ala Ala Gly Leu Val 115 120 125 Ala Asp Met Lys Ala GlyGly Ala Pro Ala Glu Leu Val Gly Ser Tyr 130 135 140 Ala Leu Ala Ile ProVal Thr Val Ile Cys Glu Leu Leu Gly Val Pro 145 150 155 160 Glu Asp AspArg Thr Arg Leu Arg Gly Trp Cys Asp Ala Ala Leu Ser 165 170 175 Thr GlyGlu Leu Thr Asp Glu Glu Cys Val Gln Ser Phe Met Asp Leu 180 185 190 GlnLys Tyr Phe Glu Asp Leu Val Lys Glu Arg Arg Ala Glu Pro Arg 195 200 205Asp Asp Leu Thr Ser Ala Leu Ile Glu Ala Arg Asp Ala His Asp Arg 210 215220 Leu Ala Glu Pro Glu Leu Ile Gly Leu Cys Ile Ser Ile Leu Ile Gly 225230 235 240 Gly Phe Glu Thr Thr Ala Ser Glu Ile Ser Ser Phe Val His ValLeu 245 250 255 Gln Gln Arg Arg Glu Leu Trp Thr Arg Leu Cys Ala Asp ProGlu Ala 260 265 270 Ile Pro Ala Ala Val Glu Glu Leu Leu Arg Phe Val ProPhe Ala Ala 275 280 285 Asn Gly Ile Ser Pro Arg Tyr Ala Leu Glu Asp MetThr Val Gly Gly 290 295 300 Val Leu Val Arg Glu Gly Glu Pro Val Ile ValAsp Thr Ser Ala Val 305 310 315 320 Asn Arg Asp Gly Leu Val Phe Asp AsnAla Asp Glu Val Val Ile Asp 325 330 335 Arg Ala Asp Asn Arg His Met ValPhe Gly His Gly Ala His His Cys 340 345 350 Leu Gly Ala His Leu Ala ArgVal Glu Leu Gln Glu Ala Leu Lys Ala 355 360 365 Leu Val Glu Gly Met ProGly Leu Arg Leu Ser Gly Asp Val Glu Trp 370 375 380 Lys Ala Asp Met IleIle Arg Ala Pro Arg Val Met His Val Glu Trp 385 390 395 400 63 134 PRTStreptomyces atroolivaceus 63 Met Thr Gln Met Arg Ile Gln Ala Thr PheVal Val Asp Val Trp Asp 1 5 10 15 Gly Thr Asp Asp Glu Pro Val Asp GlyGly Pro Val Thr Gly Arg Val 20 25 30 Glu Leu Thr Lys Thr Tyr Thr Glu GlyAsp Val Lys Gly Ser Ala Thr 35 40 45 Gly His Met Val Thr Thr Gln Gly ProGly Gly Ala Ala Tyr Val Ala 50 55 60 Gln Glu Arg Val Thr Gly Ile Met GlyGly Arg Thr Gly Thr Phe Val 65 70 75 80 Leu Glu His Arg Ala Thr Gln ValPro Gly Thr Asp Pro Val Thr Trp 85 90 95 Ala Gly Ile Val Pro Gly Ser GlyThr Gly Glu Leu Ala Gly Val Ser 100 105 110 Gly Glu Gly Ser Leu Gly HisGly Thr Leu Asp Leu Gln Val Glu Phe 115 120 125 Ala Thr Asp Gly Ala Ala130 64 216 PRT Streptomyces atroolivaceus 64 Met Ala Pro Gly Ser Gly ArgAla Arg Arg Gly Asn Glu Glu Gly Thr 1 5 10 15 Arg Thr His Met Arg ThrVal Pro Ser Asp Ala Thr Asp Ala Thr Ala 20 25 30 Pro Ala Gly Gln Thr GlyPro Ala Tyr Ile Ala Phe Leu Arg Ala Val 35 40 45 Asn Val Pro Gly Arg LysVal Glu Met Ala Arg Leu Arg Glu Val Leu 50 55 60 Thr Gln Leu Gly Phe MetGly Ala Arg Thr Tyr Ile Ala Ser Gly Asn 65 70 75 80 Ala Phe Phe Thr GlyAla Gly Asn Asp Arg Ala Arg Ile Val Glu Arg 85 90 95 Val Glu Ala Gly LeuAla Glu Ala Phe Gly Phe Glu Val Pro Thr Ile 100 105 110 Leu Arg Thr ThrAsp Glu Leu Arg Thr Glu Leu Ala Ala Ser Pro Phe 115 120 125 Val Gly ArgGln Pro Ala Pro Asp Glu Arg Phe Ser Met Val Tyr Ser 130 135 140 Ser GlyPro Leu Thr Asp Gly Glu Phe Pro Leu Arg Ser Ala Lys Gly 145 150 155 160Asp Trp Glu Val Leu Gly Ala Cys Gly Ala Thr Ala Tyr Val Arg Trp 165 170175 Arg Leu Val Asn Gly Arg Pro Val Asn Pro Val Pro Ala Ile Glu Lys 180185 190 Thr Phe Gly Val Gln Ala Thr Gly Arg Phe Phe His Thr Ser Glu Lys195 200 205 Ile Leu Ala Ala Ala Gly Lys Gly 210 215 65 272 PRTStreptomyces atroolivaceus 65 Met Asp Arg Glu Gly Asn His Arg Ala ArgArg Ile Asp Arg Pro His 1 5 10 15 Ser His Pro Gly Gly Leu Arg Met ThrGly Thr Ser Val Arg Leu Phe 20 25 30 Cys Leu Pro Tyr Ala Gly Ala Ser AlaGly Val Tyr Leu Pro Trp Cys 35 40 45 Glu Arg Leu Gln Pro His Ile Ala ValThr Pro Leu Glu Leu Pro Gly 50 55 60 Arg Gly Ser Arg Met Arg Glu Glu ProArg Arg Arg Leu Glu Pro Leu 65 70 75 80 Leu His Asp Leu Val Pro Thr ValAla Arg Leu Cys Asp Arg Pro Phe 85 90 95 Ala Leu Tyr Gly His Gly Phe GlyAla Val Leu Ala Tyr Glu Ile Ala 100 105 110 Ala Arg Leu Glu Trp Asp HisGlu Leu Val Ala Glu Arg Leu Tyr Val 115 120 125 Ser Gly Tyr Pro Ala ProHis Leu Pro Pro Arg Glu Glu Pro Ile Gly 130 135 140 His Leu Pro Asp GluGlu Phe Leu Ala Arg Ala Gly His His Pro Arg 145 150 155 160 Ile His ProGly Thr Phe Gly Glu Ala Gly Leu Ala Arg Leu Gln Leu 165 170 175 Pro ValLeu Arg Ala Asp Phe Ala Leu Ala Glu Ser Tyr Glu Glu Gly 180 185 190 LeuAsp Asn Thr Val His Ala Pro Val Thr Ala Leu Ala Gly Gln Asp 195 200 205Asp Leu Thr Val Arg Gly Ala Asp Leu Gly Gly Trp Arg Asp Tyr Thr 210 215220 Arg Arg Ser Phe Arg Ala Arg Arg Leu Pro Gly Gly His Phe His Trp 225230 235 240 His Ala Asp Glu Arg Pro Leu Leu Asn Val Phe Val Asp Asp LeuIle 245 250 255 Arg Cys Asn Ala Ala His Asn Arg Phe Pro Val Ala Glu ArgThr Phe 260 265 270 66 345 PRT Streptomyces atroolivaceus 66 Met Thr GlyThr Leu Val Arg Gly Ala Ala Arg Thr Arg Pro Arg Leu 1 5 10 15 Met AlaVal His Ala His Pro Asp Asp Glu Ser Ser Lys Gly Ala Ala 20 25 30 Thr LeuAla Arg Tyr Ala Ala Glu Gly Val Glu Val Leu Val Val Thr 35 40 45 Cys ThrGly Gly Glu Arg Gly Ser Ile Leu Asn Pro Gly Tyr Pro Arg 50 55 60 Pro ArgGly Pro Val Asp Met Ser Val Leu Arg Ala Arg Glu Met Glu 65 70 75 80 ArgAla Arg Arg Ile Leu Gly Val Arg His Ser Trp Leu Gly Tyr Val 85 90 95 AspSer Gly Leu Pro Gln Gly Asp Glu Pro Leu Pro Ala Thr Cys Phe 100 105 110Ala Arg Val Ala Pro Gln Ala Ala Thr Ala Arg Leu Val Arg Leu Ile 115 120125 Arg Gln Phe Arg Pro Asp Val Val Thr Thr Tyr Asp Glu Asp Gly Gly 130135 140 Tyr Pro His Pro Asp His Val Met Ala His Arg Ile Thr Val Ala Ala145 150 155 160 Phe Thr Ala Ala Gly Asp Gly Ala Ala His Pro Arg Ala GlyArg Pro 165 170 175 Trp Gln Pro Leu Lys Leu Tyr Tyr Asn His Gly Ser HisPro Ala Arg 180 185 190 Ser Ala Ala Leu His Arg Ala Leu Arg Glu Arg GlySer Thr Ala Pro 195 200 205 Tyr Ala Pro Leu Pro Asn Glu Pro Val Arg GlnPro Asp Ile Thr Thr 210 215 220 Arg Val Pro Cys Ala Glu Tyr Phe Pro LeuArg Asp Arg Ala Leu Arg 225 230 235 240 Ala His Thr Ser Gln Val Glu ProHis Gly Val Trp Phe Ala Val Pro 245 250 255 Thr Pro Val Gln Gln Glu ValTrp Pro Thr Glu Asp Tyr Thr Leu Ala 260 265 270 Arg Ser Leu Val Arg ThrGlu Leu Pro Glu Ser Asp Leu Phe Ala Gly 275 280 285 Ile Ala Thr Ala AlaThr Val Gly Arg Arg Gly Val Gly Pro Arg Gln 290 295 300 Ile Arg Ala ArgArg Pro Glu Gly Leu Arg Val Lys Met Pro Ala Asp 305 310 315 320 Ala AlaArg Ala Ala Leu Arg Ala Ala Ser Ser Gly Arg Ser Gly Ser 325 330 335 GlySer Ala Arg Ser Arg Thr Ser Trp 340 345 67 236 PRT Streptomycesatroolivaceus 67 Met Pro Pro Pro Pro Arg Ala Thr Gly Ile Pro Asp Thr AspSer Ala 1 5 10 15 Ala Pro His Ala Ala Ser Gly Ser Ala Thr Ala Thr ThrAla Ala Pro 20 25 30 Ala Pro Pro Val Arg Arg Pro Gly Gly Arg Thr Ala ArgAsn Arg Ala 35 40 45 Gln Val Leu Asp Ala Val Arg Ser Glu Leu Val Glu HisGly Tyr Asp 50 55 60 Ala Leu Thr Val Asp Ala Val Ala Asp Arg Ala Gly ValHis Arg Thr 65 70 75 80 Thr Val Tyr Arg Arg Trp Gly Asp Val Gly Gly LeuLeu Ala Asp Val 85 90 95 Leu Asn Ala Gly Met Gly Asp Asp Trp Glu Pro AlaAsp Thr Gly Thr 100 105 110 Leu Tyr Gly Asp Leu Ala Ala Leu Asn His AspVal Gln Thr Ala Leu 115 120 125 Ser Glu Gln Pro Ser Val Thr Ala Ala LeuIle Ala Ala Ser Phe Arg 130 135 140 Ser Glu Gln Ala Ala Arg Ala Leu ArgArg Phe Trp Asp Asp Arg Tyr 145 150 155 160 Thr Arg Cys Glu Val Thr ValGlu Arg Ala Val Glu Arg Gly Asp Val 165 170 175 Pro Pro Asp Thr Ala AlaArg Pro Leu Leu Val Ala Ala Thr Ala Pro 180 185 190 Leu Tyr His Gln LeuVal Leu Leu Arg Ala Glu Pro Asp Pro Leu Leu 195 200 205 Pro Asp Glu AlaAla Arg Ser Ala Ala Leu Ala Ala Ser Ala Gly Ile 210 215 220 Phe Thr ArgSer Pro Ser Gly Arg Arg Ala Leu Ile 225 230 235 68 539 PRT Streptomycesatroolivaceus 68 Met Pro Thr Gln Ile Ser Val Arg Ser Val Thr Lys Ser PheAsp Asp 1 5 10 15 Arg Leu Val Leu Asp Ala Val Thr Cys Ser Leu Pro AlaGly Glu Arg 20 25 30 Ala Gly Ile Ile Gly Glu Asn Gly Ser Gly Lys Ser ThrLeu Leu Arg 35 40 45 Leu Leu Ala Gly Arg Asp Gln Pro Asp Gln Gly Thr ValVal Val Gln 50 55 60 Ala Asp Gly Gly Val Gly Tyr Leu Ala Gln Asp Glu GlnLeu Pro Ser 65 70 75 80 Arg Ala Thr Val Gln Asp Val Ile Asp Arg Ala LeuGly Asp Leu Arg 85 90 95 Thr Met Glu Glu Arg Met Arg Arg Leu Glu Thr SerMet Ala Asp Gly 100 105 110 Asp Glu Ser Gly Met Ala Glu Tyr Gly Asp LeuMet Thr Val Phe Glu 115 120 125 Leu Arg Gly Gly Tyr Asp Ala Asp Ala ArgVal Glu Arg Ala Leu His 130 135 140 Gly Leu Gly Leu Asp Leu Leu Asp ArgAsp Arg Thr Val Asp Gly Leu 145 150 155 160 Ser Gly Gly Glu Gln Val ArgLeu Arg Leu Ala Ala Val Leu Ala Ala 165 170 175 Gly Pro Glu Val Leu LeuLeu Asp Glu Pro Thr Asn His Leu Asp Glu 180 185 190 Asp Ala Leu Ser TrpLeu Glu Asp His Leu Arg Thr Arg Arg Gly Thr 195 200 205 Thr Val Ala ValSer His Asp Arg Met Phe Leu Glu Arg Val Ala Thr 210 215 220 Ser Leu LeuGlu Val Asp Ala Asp Ser Arg Gly Ile Val Arg Tyr Gly 225 230 235 240 AsnGly Tyr Ala Gly Tyr Leu Ala Glu Lys Ala Ala Ala Arg Arg Arg 245 250 255Ala Ala Gln Ala His Ala Gln Trp Gln Ala Asp Val Ala Gln Leu Arg 260 265270 Glu Ala Ala Ala Thr Thr Ala Arg Arg Val Ala Pro Gly Arg Ala Met 275280 285 Lys Asp Gly Asn Lys Met Ala Tyr Asp Arg Ala Ala Gly Arg Val Gln290 295 300 Gln Ser Leu Ala Ser Arg Val Arg Gln Ala Glu Glu Arg Leu ArgArg 305 310 315 320 Leu Leu Ala Asp Pro Val Pro Ser Pro Pro Glu Pro LeuHis Phe Ala 325 330 335 Pro Val Leu Arg Ala Ala Gly Val Gln Gly Pro ValLeu Asp Ala Ala 340 345 350 Gly Ile Gly Val Ala Gly Arg Leu Asp Arg ThrAsp Leu Thr Ile Ala 355 360 365 Ala Gly Glu Arg Met Leu Ile Thr Gly ProAsn Gly Ala Gly Lys Thr 370 375 380 Thr Leu Leu Arg Val Leu Ala Asp GluLeu Ala Pro Asp Thr Gly Ser 385 390 395 400 Val Thr Arg Arg Gly Arg IleGly Tyr Leu Pro Gln Glu Pro Gly Pro 405 410 415 Gly Arg Pro Gly Glu SerLeu Leu Ala Ala Phe Ala Arg Gly Arg Ser 420 425 430 Gly Gln Thr Ala GluHis Arg Glu Arg Leu Leu Ser Leu Gly Leu Phe 435 440 445 Asp Ala Glu GlnLeu Thr Ala Ser Val Arg Gly Leu Ser Thr Gly Gln 450 455 460 Arg Gln ArgLeu Ala Leu Ala Arg Leu Val Thr Gln Pro Ser Asp Val 465 470 475 480 LeuLeu Leu Asp Glu Pro Thr Asn His Leu Ser Pro Ala Leu Val Glu 485 490 495Glu Leu Glu Ala Ala Leu Ala Gly Tyr Asp Gly Ala Leu Val Val Val 500 505510 Ser His Asp Arg Arg Leu Arg Arg Arg Trp Gln Gly Ala Gln Leu Thr 515520 525 Leu Ser Thr Asn Ala Pro Gln Ala Val Arg Val 530 535 69 322 PRTStreptomyces atroolivaceus 69 Met His His Arg Pro Ser Gly Ser Glu ThrArg Thr Ser Ala Arg Thr 1 5 10 15 Asn Ser Gln Gly Val Thr His Ala SerVal Gln Arg Arg Val Gln Ala 20 25 30 Gly Val Arg Arg Gly Pro Ser Val ProSer Gly Pro Tyr Asp Val Thr 35 40 45 Thr Gly Pro Asp Asp Ala Leu Trp PheThr Leu Ile His Ser Gly Gly 50 55 60 Ile Ala Arg Leu Thr Pro Asp Gly AlaLeu Asp Arg His Ala Leu Asp 65 70 75 80 Ser Pro Ala Cys Gly Pro Ser IleIle Thr Pro Gly Pro Asp Gly Ala 85 90 95 Leu Trp Phe Thr Arg Cys Gln AspHis Arg Ile Gly Arg Met Thr Val 100 105 110 Thr Gly His Ala Thr Ser PheAla Val Ala Thr Pro Asp Ser Gly Pro 115 120 125 Tyr Gly Ile Ala Ala GlyGly Asp Gly Ala Leu Trp Phe Thr Gln Leu 130 135 140 His Thr Asp Arg IleGly Arg Ile Thr Val Asp Gly Arg Val Thr Glu 145 150 155 160 Phe Pro LeuPro Phe Arg Gly Ala Phe Pro Ser Ala Ile Thr Ala Gly 165 170 175 Pro AspGln Ala Leu Trp Phe Thr Leu Asn Arg Ala Asp Ala Val Gly 180 185 190 ArgMet Asp Leu Gly Gly Glu Val Thr Val His Pro Leu Pro Thr Glu 195 200 205His Ala Ala Pro Val Gly Ile Thr Val Gly Thr Asp Gly Ala Leu Trp 210 215220 Phe Val Glu Ile Gly Ala Gly Gln Val Gly Arg Val Gly Thr Asp Gly 225230 235 240 Val Val Arg Glu Phe Pro Leu Pro Asp Arg Ala Cys Arg Pro HisAla 245 250 255 Ile Thr Ala Asp Pro Ala Arg Gly Val Cys Trp Phe Thr GluTrp Ala 260 265 270 Ala Asp Arg Ile Gly Ser Ile Thr Pro Asp Gly His IleGlu Glu Tyr 275 280 285 Glu Leu Pro Asp Arg Gly Ala Glu Pro His Gly LeuThr Val Ala Pro 290 295 300 Asp Gly Ala Leu Tyr Val Ala Leu Glu Thr GlyArg Ile Leu Arg Met 305 310 315 320 Glu Leu 70 551 PRT Streptomycesatroolivaceus 70 Met Thr Thr His Pro Asn Phe Ser Gly Pro Thr Ser Ile ThrCys Ala 1 5 10 15 Gly Leu Thr Tyr Ala Trp Pro Asp Gly Thr Pro Val PheGlu Asp Leu 20 25 30 Gln Val Ala Val Gly Pro Gly Arg Thr Gly Leu Ile GlyLeu Asn Gly 35 40 45 Ser Gly Lys Ser Thr Leu Leu Lys Leu Ile Ala Gly GluLeu Thr Pro 50 55 60 Ala Ser Gly Thr Val Arg Val Thr Gly Glu Val Gly TyrLeu Pro Gln 65 70 75 80 Asn Val Thr Leu Asp Thr Ser Leu Arg Val Asp ValAla Leu Gly Ile 85 90 95 Ala Glu Ala Arg Ala Ala Leu His Ala Ile Glu AlaGly Asp Ala Ser 100 105 110 Glu Glu His Phe Thr Ala Val Gly Asp Asp TrpAsp Val Glu Glu Arg 115 120 125 Ala Val Ala Met Leu Ser Gly Leu Gly LeuGly His Ile Gly Leu Asp 130 135 140 Arg Thr Ile Gly Glu Val Ser Gly GlyGlu Ser Val Leu Leu Arg Leu 145 150 155 160 Ala Ala Leu Leu Leu Ser ArgPro Asp Val Leu Leu Leu Asp Glu Pro 165 170 175 Thr Asn Asn Leu Asp LeuArg Ala Arg Arg Arg Leu Tyr Ala Thr Val 180 185 190 Glu Ala Trp Ser GlyVal Leu Val Leu Val Ser His Asp Arg Glu Leu 195 200 205 Leu Glu Arg ValAsp Gln Ile Ala Asp Leu Arg Gly Gly Glu Val Thr 210 215 220 Trp Tyr GlyGly Asn Leu Ser Ala Tyr Glu Glu Ala Leu Ala Ala Glu 225 230 235 240 GlnGlu Ala Ala Gln Arg Met Val Arg Val Ala Glu Ser Asp Leu Lys 245 250 255Lys Gln Lys Arg Glu Leu Ala Asp Ala Gln Val Lys Leu Ala Arg Arg 260 265270 Lys Arg Tyr Gly Gln Lys Met Trp Asp Thr Lys Arg Glu Pro Lys Val 275280 285 Val Met Gly Glu Arg Lys Arg Gln Ala Gln Val Ala Ala Gly Lys His290 295 300 Arg Ile Met His Glu Glu Lys Leu Ala Glu Ala Lys Asp Arg LeuAsp 305 310 315 320 Glu Ala Val Asp Ala Val Arg Asp Asp Asp Glu Ile HisVal Asp Leu 325 330 335 Pro Tyr Thr Ala Val Pro Pro Gly Arg Thr Val LeuThr Leu Arg Glu 340 345 350 Leu His Leu Ala His Gly Ala Arg Val Gln GlyGlu Phe Glu Leu Arg 355 360 365 Gly Pro Glu Arg Ile Ala Leu Val Gly ArgAsn Gly Ala Gly Lys Thr 370 375 380 Thr Leu Leu Arg Thr Ile Thr Gly GluLeu Glu Pro Val Glu Gly Glu 385 390 395 400 Ala Val Ala His Val Pro LeuArg Phe Leu Pro Gln Arg Leu Asp Val 405 410 415 Leu Asp Glu Ala Leu SerVal Ala Glu Asn Val Ala Arg Phe Ala Pro 420 425 430 Gln Ala Thr Asn AsnArg Ile Arg Ser Arg Leu Ala Arg Phe Leu Phe 435 440 445 Lys Gly Ala ArgAla Asp Gln Gln Ala Ala Thr Leu Ser Gly Gly Glu 450 455 460 Arg Phe ArgAla Thr Leu Ala Ala Leu Met Leu Ala Glu Pro Ala Pro 465 470 475 480 GlnLeu Leu Met Leu Asp Glu Pro Thr Asn Asn Leu Asp Met Ala Ser 485 490 495Val Arg Gln Leu Thr Ser Ala Leu Ser Ser Tyr Glu Gly Ala Leu Ile 500 505510 Val Ala Ser His Asp Val Pro Phe Leu Glu Ser Ile Gly Ile Thr Arg 515520 525 Trp Val Leu Leu Asp Gly Glu Leu Arg Asp Thr Thr Pro Glu Glu Val530 535 540 Arg Gly Glu Ala Val Thr Pro 545 550 71 469 PRT Streptomycesatroolivaceus 71 Met Pro Ser Gln Pro Pro Ala Asp Phe Thr Asp Phe Lys ValAla Asp 1 5 10 15 Leu Ser Leu Ala Ala Phe Gly Arg Lys Glu Ile Thr LeuAla Glu His 20 25 30 Glu Met Pro Gly Leu Met Ser Ile Arg Glu Glu Tyr AlaAla Ala Gln 35 40 45 Pro Leu Ala Gly Ala Arg Ile Thr Gly Ser Leu His MetThr Val Gln 50 55 60 Thr Ala Val Leu Ile Glu Thr Leu Val Ala Leu Gly AlaAsp Val Arg 65 70 75 80 Trp Val Ser Cys Asn Ile Tyr Ser Thr Gln Asp HisAla Ala Ala Ala 85 90 95 Ile Ala Ala Ala Gly Ile Pro Val Phe Ala Trp LysGly Glu Thr Leu 100 105 110 Glu Glu Tyr Trp Trp Cys Thr Glu Gln Ala LeuThr Trp Pro Gly His 115 120 125 Thr Gly Pro Asn Met Ile Leu Asp Asp GlyGly Asp Ala Thr Leu Leu 130 135 140 Val His Lys Gly Val Glu Tyr Arg LysThr Gly Ile Leu Pro Glu Ala 145 150 155 160 Glu Asn Glu Glu Leu Ala ValVal Arg Ala Leu Leu Asp Arg Ser Gly 165 170 175 Leu Asp Trp Thr Ala MetSer Ser Glu Ile Arg Gly Val Thr Glu Glu 180 185 190 Thr Thr Thr Gly ValHis Arg Leu Tyr Glu Met His Arg Asp Gly Thr 195 200 205 Leu Leu Phe ProAla Ile Asn Val Asn Asp Ala Val Thr Lys Ser Lys 210 215 220 Phe Asp AsnLys Tyr Gly Cys Arg His Ser Leu Ile Asp Gly Ile Asn 225 230 235 240 ArgAla Thr Asp Val Leu Ile Gly Gly Lys Thr Ala Val Val Cys Gly 245 250 255Tyr Gly Asp Val Gly Lys Gly Cys Ala Glu Ser Leu Arg Gly Gln Gly 260 265270 Ala Arg Val Ile Ile Thr Glu Ile Asp Pro Ile Cys Ala Leu Gln Ala 275280 285 Ala Met Asp Gly Tyr Gln Val Ala Thr Leu Asp Glu Val Val Asp Lys290 295 300 Ala Asp Ile Phe Val Thr Thr Thr Gly Asn Lys Asp Ile Ile MetAla 305 310 315 320 Ser Asp Met Ala Lys Met Lys His Gln Ala Ile Val GlyAsn Ile Gly 325 330 335 His Phe Asp Asn Glu Ile Asp Met Ala Gly Leu AlaGln Ile Pro Gly 340 345 350 Ile Val Lys Asp Glu Val Lys Pro Gln Val HisThr Trp Lys Phe Pro 355 360 365 Asp Gly Lys Val Leu Ile Val Leu Ser GluGly Arg Leu Leu Asn Leu 370 375 380 Gly Asn Ala Thr Gly His Pro Ser PheVal Met Ser Asn Ser Phe Ala 385 390 395 400 Asp Gln Thr Leu Ala Gln IleGlu Leu Phe Thr Lys Pro Asp Glu Tyr 405 410 415 Pro Thr Asp Val Tyr ValLeu Pro Lys His Leu Asp Glu Lys Val Ala 420 425 430 Arg Leu His Leu AspAla Leu Gly Val Lys Leu Thr Thr Leu Arg Pro 435 440 445 Glu Gln Ala SerTyr Ile Gly Val Glu Val Asp Gly Pro Tyr Lys Pro 450 455 460 Asp His TyrArg Tyr 465 72 303 PRT Streptomyces atroolivaceus 72 Met Ser Thr Leu ArgAsp Ile Leu Ala Ser Gly Thr Pro Thr Phe Ser 1 5 10 15 Phe Glu Phe PhePro Pro Arg Thr Glu Ala Gly Ser Arg Met Leu Trp 20 25 30 Gln Ala Ile ArgArg Leu Glu Ala Leu Ser Pro Thr Phe Val Ser Val 35 40 45 Thr Tyr Gly AlaGly Gly Ser Ser Arg Asp Arg Thr Val Glu Val Thr 50 55 60 Lys Arg Ile AlaAla Gln Thr Thr Leu Arg Pro Val Ala His Leu Thr 65 70 75 80 Ala Val GlyHis Ser Val Ala Glu Leu Arg His Ile Ile Gly Gln Tyr 85 90 95 Ala Asp AlaGly Val Arg Asp Val Leu Val Leu Arg Gly Asp Pro Pro 100 105 110 Gly AspPro Asn Ala Pro Trp Thr Pro His Pro Gln Gly Phe Thr His 115 120 125 AlaHis Glu Leu Val Arg Leu Val Arg Asn Leu Gly Gly Phe Ser Val 130 135 140Gly Val Ala Ala Phe Pro Glu Arg His Pro Arg Ser Ala Asp Trp Asp 145 150155 160 Ser Asp Ile Ser His Phe Val Ala Lys Cys Arg Ala Gly Ala Asp Tyr165 170 175 Ala Ile Thr Gln Met Phe Phe Asp Val Glu Asp Tyr Leu Arg LeuArg 180 185 190 Asp Arg Val Ala Ala Ala Gly Cys Asp Thr Pro Ile Ile ProGlu Ile 195 200 205 Met Pro Ala Thr Asp Val Arg Gln Ile Arg Arg Phe AlaGlu Leu Ser 210 215 220 Asn Ala Arg Phe Pro Ala Ala Leu Ala Asp Arg LeuGlu Ala Ala Ala 225 230 235 240 Gly Asp Pro Ala Glu Gly His Arg Ile GlyVal Glu Tyr Ala Thr Ser 245 250 255 Met Ala Glu Arg Leu Leu Ala Glu GlyAla Pro Gly Leu His Tyr Ile 260 265 270 Thr Leu Asn Arg Ser Thr Ala ThrLeu Glu Ile His Glu Asn Ile Gln 275 280 285 Ser Ile Arg Thr Ser Arg SerThr His Ala Leu Ala Ala Ala Arg 290 295 300 73 18 DNA Streptomycesatroolivaceus 73 atggagatgt ccgacacc 18 74 18 DNA Streptomycesatroolivaceus 74 ctactggccg ctgcccag 18 75 18 DNA Streptomycesatroolivaceus 75 atgagcgact tttcccgc 18 76 18 DNA Streptomycesatroolivaceus 76 tcagcgggac gcaggcgg 18 77 18 DNA Streptomycesatroolivaceus 77 ttgcccaagc ttcccatc 18 78 18 DNA Streptomycesatroolivaceus 78 tcagcgcagg ccgaaggc 18 79 18 DNA Streptomycesatroolivaceus 79 atggcggacg aacctgcg 18 80 18 DNA Streptomycesatroolivaceus 80 tcatcgttcc gtcctcct 18 81 18 DNA Streptomycesatroolivaceus 81 atgagcgcac ggcaggag 18 82 18 DNA Streptomycesatroolivaceus 82 tcaccgtgcc tcccggac 18 83 18 DNA Streptomycesatroolivaceus 83 gtgaccgacc cgaccgcc 18 84 18 DNA Streptomycesatroolivaceus 84 tcagcgcgtc ccgacgtc 18 85 18 DNA Streptomycesatroolivaceus 85 atggttgcga acgagggt 18 86 18 DNA Streptomycesatroolivaceus 86 tcagccgaag cggcggaa 18 87 18 DNA Streptomycesatroolivaceus 87 atgggccgcg tgtcccag 18 88 18 DNA Streptomycesatroolivaceus 88 tcagtccatg cgctgctg 18 89 18 DNA Streptomycesatroolivaceus 89 gtggcatcgc cacccacc 18 90 18 DNA Streptomycesatroolivaceus 90 tcacttgtca ccgccggt 18 91 18 DNA Streptomycesatroolivaceus 91 atgactgccg acaacctg 18 92 18 DNA Streptomycesatroolivaceus 92 tcagcccagg tagaggtc 18 93 18 DNA Streptomycesatroolivaceus 93 atgacactgg acgacctg 18 94 18 DNA Streptomycesatroolivaceus 94 tcagtcgagg ctgttggt 18 95 18 DNA Streptomycesatroolivaceus 95 ttgaccacgc tcacgttc 18 96 18 DNA Streptomycesatroolivaceus 96 tcaccgcacg aacgcgtt 18 97 18 DNA Streptomycesatroolivaceus 97 atccaattca gctcgcga 18 98 18 DNA Streptomycesatroolivaceus 98 tcaggagccc gcggccac 18 99 18 DNA Streptomycesatroolivaceus 99 atgagcgatc gggacagt 18 100 18 DNA Streptomycesatroolivaceus 100 tcagctgcgg cccgcctg 18 101 18 DNA Streptomycesatroolivaceus 101 atgcagaccc agctctcc 18 102 18 DNA Streptomycesatroolivaceus 102 tcagcggcgc tgcgcgcc 18 103 18 DNA Streptomycesatroolivaceus 103 atgaccattg aggtgcac 18 104 18 DNA Streptomycesatroolivaceus 104 tcatgcgggc acctcgcc 18 105 18 DNA Streptomycesatroolivaceus 105 atgacgctca ccgacctg 18 106 18 DNA Streptomycesatroolivaceus 106 tcatcggccg gccggcag 18 107 18 DNA Streptomycesatroolivaceus 107 atgctgctgc gccccacc 18 108 18 DNA Streptomycesatroolivaceus 108 tcagccggcc ggggccga 18 109 18 DNA Streptomycesatroolivaceus 109 atgacgcaga ccgccccc 18 110 18 DNA Streptomycesatroolivaceus 110 tcacgtccac ggcgtgct 18 111 18 DNA Streptomycesatroolivaceus 111 atgagacccg acatgagt 18 112 18 DNA Streptomycesatroolivaceus 112 tcacagaccc tcggggat 18 113 18 DNA Streptomycesatroolivaceus 113 atggccgaca cccgtacc 18 114 18 DNA Streptomycesatroolivaceus 114 tcagagcacg tatcggcg 18 115 18 DNA Streptomycesatroolivaceus 115 gtggcgccgc gcacgccg 18 116 18 DNA Streptomycesatroolivaceus 116 tcaggtccgt tccggtgc 18 117 18 DNA Streptomycesatroolivaceus 117 atgaccgaga ccctgccc 18 118 18 DNA Streptomycesatroolivaceus 118 tcagccctcc agcttctg 18 119 18 DNA Streptomycesatroolivaceus 119 atgcgatccg tccgcacc 18 120 18 DNA Streptomycesatroolivaceus 120 tcatcgctgt ccctccgc 18 121 18 DNA Streptomycesatroolivaceus 121 atgacggccg acgattcg 18 122 18 DNA Streptomycesatroolivaceus 122 tcaggcgggc gcctgttc 18 123 18 DNA Streptomycesatroolivaceus 123 atgttgagtg cggcggtt 18 124 18 DNA Streptomycesatroolivaceus 124 tcatgacggc gtcccggc 18 125 18 DNA Streptomycesatroolivaceus 125 atgagcggac ggctcacg 18 126 18 DNA Streptomycesatroolivaceus 126 tcaacgggcg ctgtagcc 18 127 18 DNA Streptomycesatroolivaceus 127 gtgtcgtcca actcccct 18 128 18 DNA Streptomycesatroolivaceus 128 tcaggccgtc ctcgccgc 18 129 18 DNA Streptomycesatroolivaceus 129 atgagcaagg tcgcggtc 18 130 18 DNA Streptomycesatroolivaceus 130 tcagggggtg cggaacac 18 131 18 DNA Streptomycesatroolivaceus 131 ttgccgacgg ccacgacg 18 132 17 DNA Streptomycesatroolivaceus 132 cagcacaggc ggggaag 17 133 18 DNA Streptomycesatroolivaceus 133 atggcagaga ccgagcag 18 134 18 DNA Streptomycesatroolivaceus 134 tcagcggcat tcgttcgt 18 135 18 DNA Streptomycesatroolivaceus 135 atgcggaccg cagatctg 18 136 18 DNA Streptomycesatroolivaceus 136 tcaccgggac gcctctgt 18 137 18 DNA Streptomycesatroolivaceus 137 gtgagccggc cggtcatc 18 138 18 DNA Streptomycesatroolivaceus 138 tcagacggat gccgctgt 18 139 18 DNA Streptomycesatroolivaceus 139 gtgcacactc acgtccgt 18 140 18 DNA Streptomycesatroolivaceus 140 tcagccttgc tgctgcag 18 141 18 DNA Streptomycesatroolivaceus 141 atggccgtga cactcaag 18 142 18 DNA Streptomycesatroolivaceus 142 tcaactcacc gccggctg 18 143 18 DNA Streptomycesatroolivaceus 143 atgtcggcta cgaggcgg 18 144 18 DNA Streptomycesatroolivaceus 144 tcaccatgcg atcggcag 18 145 18 DNA Streptomycesatroolivaceus 145 atggcacggg agcagaac 18 146 18 DNA Streptomycesatroolivaceus 146 tcacgacagg tcgagcac 18 147 18 DNA Streptomycesatroolivaceus 147 atgaagttcg cgatcgtc 18 148 18 DNA Streptomycesatroolivaceus 148 ttactcggcc acccacag 18 149 18 DNA Streptomycesatroolivaceus 149 atgacggacg gcgcgata 18 150 18 DNA Streptomycesatroolivaceus 150 tcaccgtgcg gcgccgct 18 151 18 DNA Streptomycesatroolivaceus 151 atgaccgacg cggcgagc 18 152 18 DNA Streptomycesatroolivaceus 152 tcagaaccag gcgggcgc 18 153 18 DNA Streptomycesatroolivaceus 153 gtgacggcca tcggcccg 18 154 18 DNA Streptomycesatroolivaceus 154 tcagggccgc ggcttctc 18 155 18 DNA Streptomycesatroolivaceus 155 atggtggcac tggttttc 18 156 18 DNA Streptomycesatroolivaceus 156 tcagcggcgg gcgaggac 18 157 18 DNA Streptomycesatroolivaceus 157 atgaccaccc tgaccttc 18 158 18 DNA Streptomycesatroolivaceus 158 ctagcgggcg tccggcac 18 159 18 DNA Streptomycesatroolivaceus 159 atgaccaccc tgaccttc 18 160 18 DNA Streptomycesatroolivaceus 160 tcaccacttc cgtccttc 18 161 18 DNA Streptomycesatroolivaceus 161 gtgaacgtgc cctccgca 18 162 18 DNA Streptomycesatroolivaceus 162 tcatgccggg tgctcctc 18 163 18 DNA Streptomycesatroolivaceus 163 atgaccatca cctcgtcg 18 164 18 DNA Streptomycesatroolivaceus 164 tcatgcttcc cccttcgg 18 165 18 DNA Streptomycesatroolivaceus 165 atgacccagg caccactg 18 166 18 DNA Streptomycesatroolivaceus 166 tcatcgcggg gctccgct 18 167 18 DNA Streptomycesatroolivaceus 167 atgaccgcga ccggtgcc 18 168 18 DNA Streptomycesatroolivaceus 168 tcagcgccac gcgtactg 18 169 18 DNA Streptomycesatroolivaceus 169 gtgtacggct ctcggacg 18 170 18 DNA Streptomycesatroolivaceus 170 tcacgtggca actttatg 18 171 18 DNA Streptomycesatroolivaceus 171 atgaacctgc tggatgtc 18 172 18 DNA Streptomycesatroolivaceus 172 tcagacgcat cggctctc 18 173 18 DNA Streptomycesatroolivaceus 173 atgtgggacc acaagttc 18 174 18 DNA Streptomycesatroolivaceus 174 tcatcggccg gctccgtc 18 175 18 DNA Streptomycesatroolivaceus 175 atgagcggcg ccaagctg 18 176 18 DNA Streptomycesatroolivaceus 176 tcaggacgcc ggggcgag 18 177 18 DNA Streptomycesatroolivaceus 177 ttgagcgcag tcttcgac 18 178 18 DNA Streptomycesatroolivaceus 178 tcagaccccg tcgactgc 18 179 18 DNA Streptomycesatroolivaceus 179 atgacggccc cgacgccg 18 180 18 DNA Streptomycesatroolivaceus 180 tcaaggcacg aacctcgc 18 181 18 DNA Streptomycesatroolivaceus 181 gtgacgtccg ccgtccgg 18 182 18 DNA Streptomycesatroolivaceus 182 tcatgtcgcc gtcctcat 18 183 18 DNA Streptomycesatroolivaceus 183 atgtcacggg tcaacggc 18 184 18 DNA Streptomycesatroolivaceus 184 tcacgcggac ctggcccg 18 185 17 DNA Streptomycesatroolivaceus 185 atgagcaccg acaggag 17 186 18 DNA Streptomycesatroolivaceus 186 tcaggcccac cagtcgcg 18 187 18 DNA Streptomycesatroolivaceus 187 atgacggaac ggacgttc 18 188 18 DNA Streptomycesatroolivaceus 188 tcatgacggg gctcctgt 18 189 18 DNA Streptomycesatroolivaceus 189 atggccgaca cactcctc 18 190 18 DNA Streptomycesatroolivaceus 190 tcaacccact atctggaa 18 191 18 DNA Streptomycesatroolivaceus 191 atgaccgtca ggaccgac 18 192 18 DNA Streptomycesatroolivaceus 192 tcaggcggcg gcgtcggt 18 193 18 DNA Streptomycesatroolivaceus 193 gtgagcaccg aagtggaa 18 194 18 DNA Streptomycesatroolivaceus 194 tcaccactcg acgtgcat 18 195 18 DNA Streptomycesatroolivaceus 195 atgactcaga tgcggatt 18 196 18 DNA Streptomycesatroolivaceus 196 ctaggcagcc ccgtcggt 18 197 18 DNA Streptomycesatroolivaceus 197 atggcgcccg gctccggc 18 198 18 DNA Streptomycesatroolivaceus 198 tcagcccttc ccggccgc 18 199 18 DNA Streptomycesatroolivaceus 199 gtggaccgag aggggaac 18 200 18 DNA Streptomycesatroolivaceus 200 tcagaacgtc cgctcggc 18 201 18 DNA Streptomycesatroolivaceus 201 atgaccggca cgctcgtg 18 202 18 DNA Streptomycesatroolivaceus 202 tcaccaactg gtcctgct 18 203 18 DNA Streptomycesatroolivaceus 203 atgccaccgc ctccccga 18 204 18 DNA Streptomycesatroolivaceus 204 tcagatcagg gcgcgccg 18 205 18 DNA Streptomycesatroolivaceus 205 atgcctacgc agatcagc 18 206 18 DNA Streptomycesatroolivaceus 206 tcagacccgg acggcctg 18 207 18 DNA Streptomycesatroolivaceus 207 atgcaccaca ggccgtcc 18 208 18 DNA Streptomycesatroolivaceus 208 ctagagctcc atgcgcag 18 209 18 DNA Streptomycesatroolivaceus 209 atgaccaccc acccgaac 18 210 18 DNA Streptomycesatroolivaceus 210 tcacggtgtc accgcttc 18 211 18 DNA Streptomycesatroolivaceus 211 atgccctcgc agccgccc 18 212 18 DNA Streptomycesatroolivaceus 212 tcagtagcgg tagtggtc 18 213 18 DNA Streptomycesatroolivaceus 213 ttgagcacgc tgcgcgac 18 214 18 DNA Streptomycesatroolivaceus 214 tcagcgggcg gctgcgag 18

What is claimed is:
 1. A method of loading an extender molecule on anacyl carrier protein (ACP) domain comprising the step of contacting saidACP domain with the extender molecule and a recombinantly expressed,discrete acyl transferase (AT), whereby the extender molecule is loadedonto the ACP domain by the catalytic activity of the discrete AT,provided in trans.
 2. A method according to claim 1 wherein the methodis carried out in vitro.
 3. A method according to claim 1 wherein themethod is carried out in a host cell.
 4. A method according to claim 3wherein the host cell includes a vector comprising a nucleic acidencoding the discrete AT.
 5. A method according to claim 4 wherein thehost cell includes a different vector comprising a nucleic acid encodingthe ACP domain.
 6. A method according to claim 3 wherein the host cellis a bacterium.
 7. A method according to claim 1 wherein ACP is aholo-ACP and the method includes the further step of converting anapo-acyl carrier protein (apo-ACP) to said holo-ACP.
 8. A methodaccording to claim 7 wherein the conversion is catalyzed by aphosphopantetheinyl transferase.
 9. A method according to claim 8wherein the phosphopantetheinyl transferase is Svp phosphopantetheinyltransferase.
 10. A method according to claim 1 wherein the extendermolecule is selected from the group consisting of malonyl CoA, alkylmalonyl CoA, acyl malonyl CoA, hydroxy malonyl CoA and alkoxy malonylCoA.
 11. A method according to claim 1 wherein ACP domain is selectedfrom the group consisting of lnmI-ACP3, lmnJ-ACP4, lnmJ-ACP5,lnmJ-ACP6-1, lnmJ-ACP6-2, lnmJ-ACP7 and lnmJ-ACP8.
 12. A methodaccording to claim 1 wherein the ACP domain is present within modularstructure of a type I PKS or type I PKS/NRPS.
 13. A method according toclaim 1 wherein the discrete AT provided in trans comprises apolypeptide selected from the group consisting of: (a) a catalyticdomain encoded by SEQ ID NO:43; (b) a catalytic domain encoded by anucleic acid having the sequence of a nucleic acid amplified by PCRusing the primer pair set forth in SEQ ID NO:155 and 156, respectively,using the nucleic acid of a leinamycin-producing organism as template;and (c) a catalytic domain encoded by a nucleic acid that specificallyhybridizes to SEQ ID NO:43 under stringent conditions.
 14. A methodaccording to claim 1 wherein the ACP domain is present within a type IPKS module lacking a naturally-occurring acyl transferase domain orincludes an acyl transferase domain which has been unnaturally renderedineffective for loading the extender molecule to said ACP domain.
 15. Amethod according to claim 1 wherein the AT domain is selected from thegroup of polypeptides consisting of PksC-AT, PksD-AT, PksE-AT (fromBacillus subtilis), PedC-AT, PedD-AT (from a bacterial symbiont ofPaederus beetles), MmpIII-AT1, MmpIII-AT2 (from Pseudomonasfluorescens), FenF-AT (from Bacillus subtilis), Mx-TaK-AT (fromMyxococcus xanthus) and LmnG-AT (from Streptomyces atroolivaceus).
 16. Amethod according to claim 1 wherein the discrete AT is an LnmG homolog.17. A method of loading an extender molecule on a type I polyketidesynthase (PKS) module, comprising the step of contacting said type I PKSmodule with the extender molecule and a recombinantly expressed discreteacyl transferase (AT), whereby the extender molecule is loaded onto thetype I PKS module by the acyl transferase catalytic activity of thediscrete AT, provided in trans.
 18. A method according to claim 17wherein the type I PKS module lacks a naturally-occurring acyltransferase domain or the type I PKS module includes an acyl transferasedomain which has been unnaturally rendered ineffective for loading theextender molecule to said type I PKS module.
 19. A method according toclaim 17 wherein the method is carried out in vitro.
 20. A methodaccording to claim 17 wherein the method is carried out in a host cell.21. A method according to claim 20 wherein the host cell includes avector comprising a nucleic acid encoding the discrete AT.
 22. A methodaccording to claim 21 wherein the host cell includes a different vectorcomprising a nucleic acid encoding the ACP domain.
 23. A methodaccording to claim 20 wherein the host cell is a bacterium.
 24. A methodaccording to claim 20 wherein the extender molecule is endogenouslysupplied.
 25. A method according to claim 17 wherein the extendermolecule is malonyl CoA.
 26. A method according to claim 17 wherein theextender molecule is selected from the group consisting of malonyl CoA,alkyl malonyl CoA, acyl malonyl CoA, hydroxy malonyl CoA and alkoxymalonyl CoA.
 27. A method according to claim 17 wherein the type I PKSmodule is a leinamycin biosynthesis PKS module.
 28. A method accordingto claim 17 wherein the discrete AT provided in trans comprises apolypeptide selected from the group consisting of: (a) an acyltransferase catalytic domain encoded by SEQ ID NO:43; (b) anacyltransferase catalytic domain encoded by a nucleic acid having thesequence of a nucleic acid amplified by PCR using the primer pair setforth in SEQ ID NO:155 and 156, respectively, using the nucleic acid ofa leinamycin-producing organism as template; and (c) an acyl transferasecatalytic domain encoded by a nucleic acid that specifically hybridizesto SEQ ID NO:43 under stringent conditions.
 29. A method according toclaim 17 wherein the discrete AT provided in trans is an LnmG homolog.30. A method for producing a polyketide or polyketide/nonribosomalpeptide hybrid or analog thereof comprising the steps of: (a) providinga type I polyketide synthase (PKS) or type I PKS/nonribosomal proteinsynthetase (PKS/NRPS) including a module lacking an acyltransferase (AT)domain or including an acyltransferase domain which has been unnaturallyrendered ineffective for loading the extender molecule on said module;(b) contacting the PKS or NRPS of (a) with a recombinantly-expressed,discrete acyltransferase (AT) wherein the discrete AT is catalyticallyactive in loading an extender molecule onto the module lacking in acyltransferase activity; (c) incubating under conditions permitting thebiosynthesis of the polyketide or polyketide/nonribosomal peptide oranalog thereof; and (d) isolating the polyketide orpolyketide/nonribosomal peptide or analog thereof.
 31. A methodaccording to claim 30 wherein steps (a), (b) and (c) are carried out ina host cell.
 32. A method according to claim 31 wherein the host cellincludes a vector comprising a nucleic acid encoding the discrete AT.33. A method according to claim 32 wherein the host cell includes adifferent vector comprising a nucleic acid encoding the ACP domain. 34.A method according to claim 31 wherein the extender molecule isendogenously supplied by the host cell.
 35. A method according to claim30 wherein the extender molecule is exogenously supplied.
 36. A methodof loading an extender molecule on a type I PKS or type I PKS/NRPS acylcarrier protein (ACP) domain, comprising the step of contacting said ACPdomain with the extender molecule and a recombinantly expressed AT,whereby the extender molecule is loaded onto the ACP domain by the acyltransferase catalytic activity of the AT.
 37. A method according toclaim 36 wherein the AT comprises a polypeptide selected from the groupconsisting of: (a) an acyl transferase catalytic domain encoded by SEQID NO:43; (b) an acyltransferase catalytic domain encoded by a nucleicacid having the sequence of a nucleic acid amplified by PCR using theprimer pair set forth in SEQ ID NO:155 and 156, respectively, using thenucleic acid of a leinamycin-producing organism as template; and (c) anacyl transferase catalytic domain encoded by a nucleic acid thatspecifically hybridizes to SEQ ID NO:43 under stringent conditions. 38.A method according to claim 36 wherein the method is carried out invitro.
 39. A method according to claim 36 wherein the method is carriedout in a host cell.
 40. A method according to claim 39 wherein the hostcell includes a vector comprising a nucleic acid encoding the AT.
 41. Amethod according to claim 40 wherein the host cell includes a differentvector comprising a nucleic acid encoding the ACP domain.
 42. A methodaccording to claim 36 wherein the AT is an LnmG homolog.